Biotechnology -- Biobanking -- Process and quality requirements for establishment, maintenance and characterization of mammalian cell lines

This document specifies process and quality requirements for the biobanking of mammalian (including human) cell lines. It describes requirements for the fundamental procedures of the biobank handling cell lines, such as establishment, reception, identification, propagation, preservation, storage, quality control, and distribution of cell lines. This document can be used by organizations performing biobanking activities with mammalian cell lines used for research and development, biobank users, organizations and schemes using peer-assessment and accreditation bodies. This document does not apply to biological material intended for therapeutic use. NOTE International, national or regional regulations or requirements can also apply to specific topics covered in this document.

Biotechnologie -- Biobanking -- Exigences de processus et de qualité pour la génération, le maintien et la caractérisation des lignées cellulaires de mammifères

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Status
Published
Publication Date
19-Nov-2020
Current Stage
9092 - International Standard to be revised
Start Date
12-Jan-2021
Ref Project

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INTERNATIONAL ISO
STANDARD 21709
First edition
2020-11
Biotechnology — Biobanking —
Process and quality requirements
for establishment, maintenance and
characterization of mammalian cell
lines
Biotechnologie — Biobanking — Exigences de processus et de qualité
pour la génération, le maintien et la caractérisation des lignées
cellulaires de mammifères
Reference number
ISO 21709:2020(E)
ISO 2020
---------------------- Page: 1 ----------------------
ISO 21709:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020

All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may

be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting

on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address

below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2020 – All rights reserved
---------------------- Page: 2 ----------------------
ISO 21709:2020(E)
Contents Page

Foreword ..........................................................................................................................................................................................................................................v

Introduction ................................................................................................................................................................................................................................vi

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Requirements .......................................................................................................................................................................................................... 3

4.1 General ........................................................................................................................................................................................................... 3

4.2 Legal and ethical requirements ................................................................................................................................................ 3

4.3 Facilities ........................................................................................................................................................................................................ 4

4.3.1 General...................................................................................................................................................................................... 4

4.3.2 Cell culture facility .......................................................................................................................................................... 4

4.4 Equipment ................................................................................................................................................................................................... 4

4.4.1 General...................................................................................................................................................................................... 4

4.4.2 Equipment inspection ................................................................................................................................................. 5

4.5 Reagents........................................................................................................................................................................................................ 6

4.6 Informed consent .................................................................................................................................................................................. 6

4.6.1 General...................................................................................................................................................................................... 6

4.6.2 Procedure of obtaining informed consent ................................................................................................. 6

4.6.3 Special circumstances for informed consent ........................................................................................... 7

4.6.4 Information provided to the donor/patient or the donor's/patient’s

legally designated or nominated representative .................................................................................. 7

4.6.5 Informed consent signature ................................................................................................................................... 8

4.7 Personnel ..................................................................................................................................................................................................... 8

4.7.1 General...................................................................................................................................................................................... 8

4.7.2 Personnel competence ........................................................................................................................................... ..... 9

4.7.3 Personnel training .......................................................................................................................................................... 9

4.7.4 Biorisk and biosafety of personnel .................................................................................................................. 9

5 Process requirements ..................................................................................................................................................................................... 9

5.1 Establishment of cell lines within the biobank ........................................................................................................... 9

5.1.1 General...................................................................................................................................................................................... 9

5.1.2 Isolation and purification of primary cells .............................................................................................10

5.1.3 Primary cultures ...........................................................................................................................................................10

5.1.4 Cell lines ...............................................................................................................................................................................11

5.2 Reception of established cell lines ......................................................................................................................................11

5.2.1 General...................................................................................................................................................................................11

5.2.2 Review of the deposit request ...........................................................................................................................11

5.2.3 Decision on the deposit request ......................................................................................................................12

5.2.4 Transfer of material and associated data ................................................................................................12

5.2.5 Characterization and authentication of the cell line ......................................................................13

5.2.6 Assignment of accession number ...................................................................................................................13

5.3 Cell line management .....................................................................................................................................................................14

5.3.1 Planning for cell banking .......................................................................................................................................14

5.3.2 Accession .............................................................................................................................................................................14

5.3.3 Propagation of cell lines .........................................................................................................................................14

5.3.4 Preservation and storage .......................................................................................................................................15

5.3.5 Inventory management ...........................................................................................................................................15

5.3.6 Disposal management ..............................................................................................................................................15

5.4 Distribution .............................................................................................................................................................................................16

5.4.1 General...................................................................................................................................................................................16

5.4.2 Acceptance of order request ...............................................................................................................................16

5.4.3 Distribution review ....................................................................................................................................................16

5.4.4 Transport .............................................................................................................................................................................16

© ISO 2020 – All rights reserved iii
---------------------- Page: 3 ----------------------
ISO 21709:2020(E)

5.5 Quality control, validation and verification ................................................................................................................17

5.5.1 Quality control ................................................................................................................................................................17

5.5.2 Validation and verification ...................................................................................................................................17

6 Performance evaluation and improvement ..........................................................................................................................17

Annex A (informative) Recommended units for biobanking of cell lines ...................................................................19

Bibliography .............................................................................................................................................................................................................................20

iv © ISO 2020 – All rights reserved
---------------------- Page: 4 ----------------------
ISO 21709:2020(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www .iso .org/ patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see www .iso .org/

iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 276, Biotechnology.

Any feedback or questions on this document should be directed to the user’s national standards body. A

complete listing of these bodies can be found at www .iso .org/ members .html.
© ISO 2020 – All rights reserved v
---------------------- Page: 5 ----------------------
ISO 21709:2020(E)
Introduction

Scientific research using cell lines has contributed greatly to the understanding of human health. Cell

cultures are increasingly used to complement studies using animal models. Although cell lines are

important research tools, potential problems have recently been identified.

Cell lines have unique characteristics and behaviour that can change as they continue to be passaged.

The original phenotype (e.g. expression of specific biomarkers) can be lost or new characteristics or

behaviour (e.g. development of tumorigenicity) may develop. It is important to minimize passaging to

retain the original characteristics that were present when the cell line was first established.

Other problems such as contamination, either with microorganisms or another cell line, and

misidentification can also arise. Cultures can become contaminated during cell line establishment or

later when cultures are passaged. These problems are often not visible by eye and require specific

testing to be detected.

In order to help address these issues, the research community has called for an international effort

to create standards for biobanks. ISO 20387 was published to provide an overarching standard

for biobanks. This document provides additional technical specifications for biobanks that handle

mammalian cell lines. Such biobanks can demonstrate their competence in biobanking by complying

with the specifications within this document, in addition to the requirements prescribed in ISO 20387.

In this document, the following verbal forms are used:
— “shall” indicates a requirement;
— “should” indicates a recommendation;
— “may” indicates a permission;
— “can” indicates a possibility or a capability.
Further details can be found in the ISO/IEC Directives, Part 2.
vi © ISO 2020 – All rights reserved
---------------------- Page: 6 ----------------------
INTERNATIONAL STANDARD ISO 21709:2020(E)
Biotechnology — Biobanking — Process and quality
requirements for establishment, maintenance and
characterization of mammalian cell lines
1 Scope

This document specifies process and quality requirements for the biobanking of mammalian (including

human) cell lines. It describes requirements for the fundamental procedures of the biobank handling

cell lines, such as establishment, reception, identification, propagation, preservation, storage, quality

control, and distribution of cell lines.

This document can be used by organizations performing biobanking activities with mammalian

cell lines used for research and development, biobank users, organizations and schemes using peer-

assessment and accreditation bodies.

This document does not apply to biological material intended for therapeutic use.

NOTE International, national or regional regulations or requirements can also apply to specific topics

covered in this document.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 20387:2018, Biotechnology — Biobanking — General requirements for biobanking

ISO 20391-1, Biotechnology — Cell counting — Part 1: General guidance on cell counting methods

ISO 20391-2, Biotechnology — Cell counting — Part 2: Experimental design and statistical analysis to

quantify counting method performance
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1
cell culture

growth of cells dissociated from the parent tissue by spontaneous migration or mechanical or enzymatic

dispersal for propagation and consecutive passages in vitro
Note 1 to entry: Additional information can be found in Reference [6].
© ISO 2020 – All rights reserved 1
---------------------- Page: 7 ----------------------
ISO 21709:2020(E)
3.2
cell line

progeny of a primary culture (3.16) after it has been passaged beyond crises or beyond senescence

either spontaneously or after introduction of immortalizing factors
Note 1 to entry: A cell line is continuous for proliferation.
Note 2 to entry: Additional information can be found in Reference [7].
3.3
cell morphology
form and structure of the cell

Note 1 to entry: Morphology can be represented by a single parameter or a combination of two or more

parameters.
3.4
cell strain

progeny of a primary culture (3.16) before it has been passaged beyond crises or beyond senescence or

immortalized by introduction of immortalizing factors

Note 1 to entry: Not all the cell strains will continue proliferation to reach the cell lines.

3.5
cell type
classification used to distinguish among distinct cell forms
3.6
cryopreservation

process by which cells are maintained in a low temperature (e.g. −70 °C to −80 °C) at an inactive state so

they can be revived later
3.7
derivative material
biological material that was derived from the original donation
EXAMPLE Derivative materials can be cell lines.
3.8
doubling time
time taken for cultured cell count to double
3.9
establishment of cell line
process of producing a cell line capable of indefinite proliferation
3.10
identity verification

part of the process of verifying authenticity of a cell line in which cell origin is genetically confirmed

3.11
informed consent

process by which an individual or its designated legal representative or nominated representative

voluntarily confirms willingness to donate biological material for research purposes, after having been

informed of all aspects of the potential research that are relevant for the decision to donate

3.12
microbe
microorganism

unicellular living cells which cannot be observed with naked eye but only under a microscope

2 © ISO 2020 – All rights reserved
---------------------- Page: 8 ----------------------
ISO 21709:2020(E)
3.13
passage number
number of subculturing that occurred
3.14
population doubling level
PDL

total number of population doublings of a cell line (3.2) or cell strain (3.4) since its initiation in vitro

Note 1 to entry: Additional information can be found in Reference [7].
3.15
primary cells

cells isolated directly from tissue or organs taken directly from an organism, using enzymatic or

mechanical methods
3.16
primary culture

culture started from cells, tissues, or organs taken directly from an organism, and before the first

subculture, propagation and consecutive passages in vitro
Note 1 to entry: Additional information can be found in Reference [6].
3.17
viability

attribute of being alive (e.g. metabolically active, capable of reproducing, have intact cell membrane, or

have the capacity to resume these functions) as defined based on the intended use

4 Requirements
4.1 General

During the whole cell culture workflow, precautions shall be taken to avoid cross contamination

between different samples/specimens, e.g. by using disposable material whenever feasible or

appropriate cleaning procedures between processing of different specimens/samples.

The biobank shall establish, implement, and maintain a quality management system as described in

ISO 20387:2018, Clause 8.
4.2 Legal and ethical requirements
ISO 20387:2018, 4.3 shall be followed.

The biobank shall consult an ethics review committee, which is responsible for the investigation and

evaluation of any related ethical principles/requirements.

The biobank shall comply with ISO 20387:2018, 4.1.6. The biological material, which is the source of the

cell line, shall be collected, transported, and handled according to internationally accepted procedures.

The biobank shall be aware of and able to demonstrate compliance with relevant national, regional

and international approved ethics, laws and regulations relating to the biological material held in the

biobank.

Further ethical requirements related to the informed consent are included in 4.6.

The biobank shall meet the organizational structure, resource, process, and quality requirements

described in ISO 20387:2018, Clauses 5, 6 and 7, in addition to those in this document.

© ISO 2020 – All rights reserved 3
---------------------- Page: 9 ----------------------
ISO 21709:2020(E)
4.3 Facilities
4.3.1 General
ISO 20387:2018, 6.3 shall be followed.

The processing facility shall be constructed and operated to minimize the introduction, generation, and

retention of particles and microorganisms.

The biobank shall define a protocol to test the quality of the biobanking environment(s).

Where appropriate, test methods proposed in the ISO 14644 series can be considered.

4.3.2 Cell culture facility

The biobank culturing and preserving cell lines shall establish a cell culture facility.

The biobank shall assess the biological safety level of the cell lines that will be handled. The biobank

shall assess the risk of asphyxiation based on the number and condition of liquid nitrogen tanks and

storage methods and install an oxygen monitor accordingly.

The biobank shall meet biological safety and security requirements for its facilities to minimize the risk

to personnel and surroundings from handling the cell lines.
The biobank shall take necessary measures to control airflow.

The biobank should make efforts to secure a separate space for storage of duplicated cell lines in order

to prevent the loss of collections in emergency situations. If the biobank cannot secure a separate

alternate space for storage, the biobank should duplicate cell lines and store them in separate liquid

nitrogen tanks. If the biobank cannot duplicate all stored cell lines, the biobank should make duplicates

of the relevant cell lines or those that can be required by contractual agreements.

The biobank shall designate a separate area for handling deposited cell lines and/or tissues until they

are determined to be free of microbial contamination.
4.4 Equipment
4.4.1 General
ISO 20387:2018, 6.5 shall be followed.

Equipment used in biobanking should be selected under quality specific criteria defined by the biobank,

and for managing risks that can affect or impact performed assays, tests or their validity.

All incubators used, for working in accordance with this document, shall be monitored.

The biobank shall be furnished with or have controlled access to the following equipment as minimum

requirements for cell culturing and ensure quality and safety requirements regarding the equipment.

a) Biological safety cabinet (Class II, Class III): physical structure protecting cell cultures from

contaminants and personnel from potentially harmful biological substances. The cabinet can

require appropriate biosafety measures, depending upon the biosafety level of the biological

materials being handled.

b) Cell counting equipment: an automated cell counting feature is recommended, manual cell counting

slide (e.g. hemocytometer slide) may also be used.
c) CO incubator: a self-cleaning feature is recommended;
d) Freezer: data logger, alarming system and backup system are recommended.
4 © ISO 2020 – All rights reserved
---------------------- Page: 10 ----------------------
ISO 21709:2020(E)
e) Refrigerator: data logger, alarming system and backup system are recommended.

f) Liquid nitrogen tank for vapor/liquid phase storage of cell lines with temperature monitor and

alarming system are recommended.
g) microscope: a camera for documentation is recommended;

h) Other general laboratory equipment (e.g. autoclave, deep freezer, refrigerator, centrifuge).

i) Inverted microscope (equipped with a camera).

The biobank should be furnished with or have controlled access to the equipment used for controlled

rate freezing of cell lines.

The biobank shall equip its facility with appropriate biosafety equipment based on the assessed

biological safety level (documented risk analysis); see 4.3.2. A biobank facility that uses liquid nitrogen

shall have adequate ventilation.

The biobank shall have personal protective equipment available in accordance with the biosafety

level of the facility and the nature of the cell line. Personnel within the facility shall wear appropriate

personal protective equipment during cell culturing to prevent contact with culture media or aerosol

that can be produced during the process.

Personnel handling frozen materials or liquid nitrogen shall use cryogenic personal protective

equipment.

Storage equipment for cell lines should have a monitoring system and an alarming device or remote

control system for immediate response to equipment malfunction or breakdown. The biobank shall

keep records of equipment malfunction or breakdown and appropriate follow-up measures. The

biobank should have an emergency generator or other equipment necessary to protect cell lines when

power is lost.

The biobank should have the necessary backup equipment (e.g. vacant gas CO incubators, freezers, and

liquid nitrogen tanks) available in the case of equipment malfunction or failure.

4.4.2 Equipment inspection

The biobank shall inspect at planned intervals the following pieces of equipment, and their

characteristics, used for cell lines, when applicable:

a) autoclave: according to the safety level, pressure, temperature, safety valve, steam outlet, cleaning

and microbiological and chemical control of the process;

b) biological safety cabinet: airflow, HEPA filter condition, sterility and cleaning;

c) CO incubator: temperature, humidity, CO system, sterility and cleaning;
2 2

d) liquid nitrogen tank: liquid nitrogen level, temperature and inspection of alarming system;

e) microscope: components and cleaning;
f) refrigerator: temperature and stability of electric supply;

g) ultra-low temperature freezer: status of compressor, temperature system, and stability of

electric supply;
h) ventilation system: inspection;
i) oxygen monitor devices: inspection;

j) backup equipment: status and stability of electrical supply and/or supply of nitrogen or CO at

emergency of electricity failure.
© ISO 2020 – All rights reserved 5
---------------------- Page: 11 ----------------------
ISO 21709:2020(E)

Inspection records shall be retained. Details of any deviations shall be documented.

4.5 Reagents

Reagents in biobanking should be selected under quality specific criteria defined by the biobank, and

for managing risks that can affect or impact performed assays, tests or their validity.

The biobank should establish, document and implement a procedure for evaluating the reagent stability;

this procedure should define criteria for internal or external quality control (i.e. antibodies, diluents,

calibrators).
4.6 Informed consent
4.6.1 General

A procedure for obtaining the informed consent shall be established, documented and implemented.

This informed consent procedure applies to human donors/patients and the responsible party for donor

animals. In case of donations from animals, the donor's legally designated or nominated representative

can be understood as the animal’s responsible party.

The informed consent procedure shall comply with the requirements set by relevant ethics review

committees. Appropriate approval of the procedure shall be obtained from the relevant ethical

committee. Evidence of ethical approval shall be available.

The informed consent procedure shall take into account language barriers, condition and the

educ
...

FINAL
INTERNATIONAL ISO/FDIS
DRAFT
STANDARD 21709
ISO/TC 276
Biotechnology — Biobanking —
Secretariat: DIN
Process and quality requirements
Voting begins on:
2020­04­28 for establishment, maintenance and
characterization of mammalian cell
Voting terminates on:
2020­06­23
lines
RECIPIENTS OF THIS DRAFT ARE INVITED TO
SUBMIT, WITH THEIR COMMENTS, NOTIFICATION
OF ANY RELEVANT PATENT RIGHTS OF WHICH
THEY ARE AWARE AND TO PROVIDE SUPPOR TING
DOCUMENTATION.
IN ADDITION TO THEIR EVALUATION AS
Reference number
BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO­
ISO/FDIS 21709:2020(E)
LOGICAL, COMMERCIAL AND USER PURPOSES,
DRAFT INTERNATIONAL STANDARDS MAY ON
OCCASION HAVE TO BE CONSIDERED IN THE
LIGHT OF THEIR POTENTIAL TO BECOME STAN­
DARDS TO WHICH REFERENCE MAY BE MADE IN
NATIONAL REGULATIONS. ISO 2020
---------------------- Page: 1 ----------------------
ISO/FDIS 21709:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020

All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may

be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting

on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address

below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH­1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2020 – All rights reserved
---------------------- Page: 2 ----------------------
ISO/FDIS 21709:2020(E)
Contents Page

Foreword ..........................................................................................................................................................................................................................................v

Introduction ................................................................................................................................................................................................................................vi

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Requirements .......................................................................................................................................................................................................... 3

4.1 General ........................................................................................................................................................................................................... 3

4.2 Legal and ethical requirements ................................................................................................................................................ 3

4.3 Facilities ........................................................................................................................................................................................................ 4

4.3.1 General...................................................................................................................................................................................... 4

4.3.2 Cell culture facility .......................................................................................................................................................... 4

4.4 Equipment ................................................................................................................................................................................................... 4

4.4.1 General...................................................................................................................................................................................... 4

4.4.2 Equipment inspection ................................................................................................................................................. 5

4.5 Reagents........................................................................................................................................................................................................ 6

4.6 Informed consent .................................................................................................................................................................................. 6

4.6.1 General...................................................................................................................................................................................... 6

4.6.2 Procedure of obtaining informed consent ................................................................................................. 6

4.6.3 Special circumstances for informed consent ........................................................................................... 7

4.6.4 Information provided to the donor/patient or the donor's/patient’s

legally designated representative ..................................................................................................................... 7

4.6.5 Informed consent signature ................................................................................................................................... 8

4.7 Personnel ..................................................................................................................................................................................................... 8

4.7.1 General...................................................................................................................................................................................... 8

4.7.2 Personnel competence ........................................................................................................................................... ..... 8

4.7.3 Personnel training .......................................................................................................................................................... 9

4.7.4 Biorisk and biosafety of personnel .................................................................................................................. 9

5 Process requirements ..................................................................................................................................................................................... 9

5.1 Establishment of cell lines within the biobank ........................................................................................................... 9

5.1.1 General...................................................................................................................................................................................... 9

5.1.2 Isolation and purification of primary cells .............................................................................................10

5.1.3 Primary cultures ...........................................................................................................................................................10

5.1.4 Cell lines ...............................................................................................................................................................................10

5.2 Reception of established cell lines ......................................................................................................................................11

5.2.1 General...................................................................................................................................................................................11

5.2.2 Review of the deposit request ...........................................................................................................................11

5.2.3 Decision on the deposit request ......................................................................................................................11

5.2.4 Transfer of material and associated data ................................................................................................12

5.2.5 Characterization and authentication of the cell line ......................................................................12

5.2.6 Assignment of accession number ...................................................................................................................13

5.3 Cell line management .....................................................................................................................................................................13

5.3.1 Planning for cell banking .......................................................................................................................................13

5.3.2 Accession .............................................................................................................................................................................14

5.3.3 Propagation of cell lines .........................................................................................................................................14

5.3.4 Preservation and storage .......................................................................................................................................15

5.3.5 Inventory management ...........................................................................................................................................15

5.3.6 Disposal management ..............................................................................................................................................15

5.4 Distribution .............................................................................................................................................................................................16

5.4.1 General...................................................................................................................................................................................16

5.4.2 Acceptance of order request ...............................................................................................................................16

5.4.3 Distribution review ....................................................................................................................................................16

5.4.4 Transport .............................................................................................................................................................................16

© ISO 2020 – All rights reserved iii
---------------------- Page: 3 ----------------------
ISO/FDIS 21709:2020(E)

5.5 Quality control, validation and verification ................................................................................................................17

5.5.1 Quality control ................................................................................................................................................................17

5.5.2 Validation and verification ...................................................................................................................................17

6 Performance evaluation and improvement ..........................................................................................................................17

Annex A (informative) Recommended units for biobanking of cell lines ...................................................................18

Bibliography .............................................................................................................................................................................................................................19

iv © ISO 2020 – All rights reserved
---------------------- Page: 4 ----------------------
ISO/FDIS 21709:2020(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non­governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www .iso .org/ patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see www .iso .org/

iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 276, Biotechnology.

Any feedback or questions on this document should be directed to the user’s national standards body. A

complete listing of these bodies can be found at www .iso .org/ members .html.
© ISO 2020 – All rights reserved v
---------------------- Page: 5 ----------------------
ISO/FDIS 21709:2020(E)
Introduction

Scientific research using cell lines has contributed greatly to the understanding of human health. Cell

cultures are increasingly used to complement studies using animal models. Although cell lines are

important research tools, potential problems have recently been identified.

Cell lines have unique characteristics and behaviour that can change as they continue to be passaged.

The original phenotype (e.g. expression of specific biomarkers) can be lost or new characteristics or

behaviour (e.g. development of tumorigenicity) may develop. It is important to minimize passaging to

retain the original characteristics that were present when the cell line was first established.

Other problems such as contamination, either with microorganisms or another cell line, and

misidentification can also arise. Cultures can become contaminated during cell line establishment or

later when cultures are passaged. These problems are often not visible by eye and require specific

testing to be detected.

In order to help address these issues, the research community has called for an international effort

to create standards for biobanks. ISO 20387 was published to provide an overarching standard

for biobanks. This document provides additional technical specifications for biobanks that handle

mammalian cell lines. Such biobanks can demonstrate their competence in biobanking by complying

with the specifications within this document, in addition to the requirements prescribed in ISO 20387.

In this document, the following verbal forms are used:
— “shall” indicates a requirement;
— “should” indicates a recommendation;
— “may” indicates a permission;
— “can” indicates a possibility or a capability.
Further details can be found in the ISO/IEC Directives, Part 2.
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FINAL DRAFT INTERNATIONAL STANDARD ISO/FDIS 21709:2020(E)
Biotechnology — Biobanking — Process and quality
requirements for establishment, maintenance and
characterization of mammalian cell lines
1 Scope

This document specifies process and quality requirements for the biobanking of mammalian (including

human) cell lines. It describes requirements for the fundamental procedures of the biobank handling

cell lines, such as establishment, reception, identification, propagation, preservation, storage, quality

control, and distribution of cell lines.

This document can be used by organizations performing biobanking activities with mammalian

cell lines used for research and development, biobank users, organizations and schemes using peer­

assessment and accreditation bodies.

This document does not apply to biological material intended for therapeutic use.

NOTE International, national or regional regulations or requirements can also apply to specific topics

covered in this document.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 20387:2018, Biotechnology — Biobanking — General requirements for biobanking

ISO 20391­1, Biotechnology — Cell counting — Part 1: General guidance on cell counting methods

ISO 20391­2, Biotechnology — Cell counting — Part 2: Experimental design and statistical analysis to

quantify counting method performance
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at http:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1
cell culture

growth of cells dissociated from the parent tissue by spontaneous migration or mechanical or enzymatic

dispersal for propagation and consecutive passages in vitro
Note 1 to entry: Additional information can be found in Reference [6].
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ISO/FDIS 21709:2020(E)
3.2
cell line

progeny of a primary culture (3.16) after it has been passaged beyond crises or beyond senescence

either spontaneously or after introduction of immortalizing factors
Note 1 to entry: A cell line is continuous for proliferation.
Note 2 to entry: Additional information can be found in Reference [7].
3.3
cell morphology
form and structure of the cell

Note 1 to entry: Morphology can be represented by a single parameter or a combination of two or more

parameters.
3.4
cell strain

progeny of a primary culture (3.6) before it has been passaged beyond crises or beyond senescence or

immortalized by introduction of immortalizing factors

Note 1 to entry: Not all the cell strains will continue proliferation to reach the cell lines.

3.5
cell type
classification used to distinguish among distinct cell forms
3.6
cryopreservation

process by which cells are maintained in a low temperature (−70 °C to −80 °C) at inactive state so they

can be revived later
3.7
derivative material
biological material that was derived from the original donation
EXAMPLE Derivative materials can be cell lines.
3.8
doubling time
time taken for cultured cell count to double
3.9
establishment of cell line
process of producing a cell line capable of indefinite proliferation
3.10
identity verification

part of the process of verifying authenticity of a cell line in which cell origin is genetically confirmed

3.11
informed consent

process by which an individual or its designated legal representative voluntarily confirms willingness

to donate biological material for research purposes, after having been informed of all aspects of the

potential research that are relevant for the decision to donate
3.12
microbe
microorganism

unicellular living cells which cannot be observed with naked eye but only under a microscope

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ISO/FDIS 21709:2020(E)
3.13
passage number
number of subculturing that occurred
3.14
population doubling level
PDL

total number of population doublings of a cell line (3.2) or cell strain (3.4) since its initiation in vitro

Note 1 to entry: Additional information can be found in Reference [7].
3.15
primary cells

cells isolated directly from tissue or organs taken directly from an organism, using enzymatic or

mechanical methods
3.16
primary culture

culture started from cells, tissues, or organs taken directly from an organism, and before the first

subculture, propagation and consecutive passages in vitro
Note 1 to entry: Additional information can be found in Reference [6].
3.17
viability

attribute of being alive (e.g. metabolically active, capable of reproducing, have intact cell membrane, or

have the capacity to resume these functions) as defined based on the intended use

4 Requirements
4.1 General

During the whole cell culture workflow, precautions shall be taken to avoid cross contamination

between different samples/specimens, e.g. by using disposable material whenever feasible or

appropriate cleaning procedures between processing of different specimens/samples.

The biobank shall establish, implement, and maintain a quality management system as described in

ISO 20387:2018, Clause 8.

The biobank shall meet the organizational structure, resource, process, and quality requirements

described in ISO 20387:2018, Clauses 5, 6 and 7, in addition to those in this document.

4.2 Legal and ethical requirements

The biobank shall consult an ethics review committee, which is responsible for the investigation and

evaluation of any related ethical principles/requirements.

The biobank shall comply with ISO 20387:2018, 4.1.6. The biological material, which is the source of the

cell line, shall be collected, transported, and handled according to internationally accepted procedures.

The biobank shall be aware of and able to demonstrate compliance with relevant national, regional

and international approved ethics, laws and regulations relating to the biological material held in the

biobank.

Further ethical requirements related to the informed consent are included in 4.6.

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ISO/FDIS 21709:2020(E)
4.3 Facilities
4.3.1 General
ISO 20387:2018, 6.3 shall be followed.

The processing facility shall be constructed and operated to minimize the introduction, generation, and

retention of particles and microorganisms.

The biobank shall define a protocol to test the quality of the biobanking environment(s).

Where appropriate, test methods proposed in the ISO 14644 series can be considered.

4.3.2 Cell culture facility

The biobank culturing and preserving cell lines shall establish a cell culture facility.

The biobank shall assess the biological safety level of the cell lines that will be handled. The biobank

shall assess the risk of asphyxiation based on the number and condition of liquid nitrogen tanks and

storage methods and install an oxygen monitor accordingly.

The biobank shall meet biological safety and security requirements for its facilities to minimize the risk

to personnel and surroundings from handling the cell lines.
The biobank shall take necessary measures to control airflow.

The biobank should make efforts to secure a separate space for storage of duplicated cell lines in order

to prevent the loss of collections in emergency situations. If the biobank cannot secure a separate

alternate space for storage, the biobank should duplicate cell lines and store them in separate liquid

nitrogen tanks. If the biobank cannot duplicate all stored cell lines, the biobank should make duplicates

of the relevant cell lines or those that can be required by contractual agreements.

The biobank shall designate a separate area for handling deposited cell lines and/or tissues until they

are determined to be free of microbial contamination.
4.4 Equipment
4.4.1 General
ISO 20387:2018, 6.5 shall be followed.

Equipment used in biobanking should be selected under quality specific criteria defined by the biobank,

and for managing risks that can affect or impact performed assays, tests or their validity.

All incubators used, for working in accordance with this document, shall be monitored.

The biobank shall be furnished with or have controlled access to the following equipment as minimum

requirements for cell culturing and ensure quality and safety requirements regarding the equipment.

a) Biological safety cabinet (Class II, Class III): physical structure protecting cell cultures from

contaminants and personnel from potentially harmful biological substances. The cabinet can

require appropriate biosafety measures, depending upon the biosafety level of the biological

materials being handled.

b) Cell counting equipment: an automated cell counting feature is recommended, manual cell counting

slide (e.g. hemocytometer slide) may also be used.
c) CO incubator: a self-cleaning feature is recommended;
d) Freezer: data logger, alarming system and backup system are recommended.
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e) Refrigerator: data logger, alarming system and backup system are recommended.

f) Liquid nitrogen tank for vapor/liquid phase storage of cell lines with temperature monitor and

alarming system are recommended.
g) microscope: a camera for documentation is recommended;

h) Other general laboratory equipment (e.g. autoclave, deep freezer, refrigerator, centrifuge).

i) Inverted microscope (equipped with a camera).

The biobank should be furnished with or have controlled access to the equipment used for controlled

rate freezing of cell lines.

The biobank shall equip its facility with appropriate biosafety equipment based on the assessed

biological safety level (documented risk analysis); see 4.3.2. A biobank facility that uses liquid nitrogen

shall be equipped with an air suction device.

The biobank shall have personal protective equipment available in accordance with the biosafety

level of the facility and the nature of the cell line. Personnel within the facility shall wear appropriate

personal protective equipment during cell culturing to prevent contact with culture media or aerosol

that can be produced during the process.

Personnel handling frozen materials or liquid nitrogen shall use cryogenic personal protective

equipment.

Storage equipment for cell lines should have a monitoring system and an alarming device or remote

control system for immediate response to equipment malfunction or breakdown. The biobank shall

keep records of equipment malfunction or breakdown and appropriate follow-up measures. The

biobank should have an emergency generator or other equipment necessary to protect cell lines when

power is lost.

The biobank should have the necessary backup equipment (e.g. vacant gas CO incubators, freezers, and

liquid nitrogen tanks) available in the case of equipment malfunction or failure.

4.4.2 Equipment inspection

The biobank shall inspect at planned intervals the following pieces of equipment, and their

characteristics, used for cell lines, when applicable:

a) autoclave: according to the safety level, pressure, temperature, safety valve, steam outlet, cleaning

and microbiological and chemical control of the process;

b) biological safety cabinet: airflow, HEPA filter condition, sterility and cleaning;

c) CO incubator: temperature, humidity, CO system, sterility and cleaning;
2 2

d) liquid nitrogen tank: liquid nitrogen level, temperature and inspection of alarming system;

e) microscope: components and cleaning;
f) refrigerator: temperature and stability of electric supply;

g) ultra-low temperature freezer: status of compressor, temperature system, and stability of

electric supply;
h) ventilation system: inspection;
i) oxygen monitor devices: inspection;

j) backup equipment: status and stability of electrical supply and/or supply of nitrogen or CO at

emergency of electricity failure.
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It is required to make records of checking the above-mentioned operating parameters of the device

and to record deviations from the above-mentioned parameters. It is required to clarify the registered

deviations and to document them.
4.5 Reagents

Reagents in biobanking should be selected under quality specific criteria defined by the biobank, and

for managing risks that can affect or impact performed assays, tests or their validity.

The biobank should establish, document and implement a procedure for evaluating the reagent stability;

this procedure should define criteria for internal or external quality control (i.e. antibodies, diluents,

calibrators).
4.6 Informed consent
4.6.1 General

A procedure for obtaining the informed consent shall be established, documented and implemented.

...

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