ISO 21709:2020
(Main)Biotechnology — Biobanking — Process and quality requirements for establishment, maintenance and characterization of mammalian cell lines
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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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
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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
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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
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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].
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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
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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.
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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;
2
d) Freezer: data logger, alarming system and backup system are recommended.
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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
2
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
2
emergency of electricity failure.
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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
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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
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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.
vi © ISO 2020 – All rights reserved
---------------------- Page: 6 ----------------------
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].
© ISO 2020 – All rights reserved 1
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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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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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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;
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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
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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;
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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
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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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