ISO 24603:2022

INTERNATIONAL ISO
STANDARD 24603
First edition
2022-08
Biotechnology — Biobanking —
Requirements for human and mouse
pluripotent stem cells
Biotechnologie — Biobanking — Exigences relatives aux cellules
souches pluripotentes humaines et murines
Reference number
ISO 24603:2022(E)
© ISO 2022

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ISO 24603:2022(E)
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© ISO 2022
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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Published in Switzerland
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ISO 24603:2022(E)
Contents Page
Foreword .v
Introduction . vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Abbreviated terms . 5
5 General requirements . 6
5.1 General . 6
5.2 Legal and ethical requirements . 6
5.3 Personnel, facilities and equipment . 6
5.4 Reagents, consumables and other supplies . 7
5.5 Management of information and data . 7
6 Collection of biological source materials and associated data. 8
6.1 Information about the human donor and requirements for the biological material . 8
6.2 Information about the mouse donor and requirements for the biological material . 8
6.3 Collection procedure . 9
7 Transport of the biological source material or PSCs and associated data to the
biobank . 9
8 Reception and traceability of the biological source material or PSCs and associated
data .10
9 Establishment of cell lines .10
9.1 Processes . 10
9.2 Unique identification . 10
9.3 Testing for infectious agents . 10
10 Characterization .11
10.1 General . 11
10.2 Population doubling time and subculture/passage.12
10.2.1 PDT . 12
10.2.2 Subculture/passage .13
10.3 Stability of the culture .13
10.4 Functionality . 14
10.4.1 General . 14
10.4.2 In vitro differentiation . 14
10.5 Microbial contamination . 14
11 Quality control .14
12 Testing.15
13 Cell line management .15
14 Preservation of cell lines .15
15 Storage .16
16 Thawing .16
17 Disposal .17
18 Distribution .17
18.1 General requirements . 17
18.2 Information for users . 17
18.3 Transport . 18
Annex A (informative) Potency comparison .21
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ISO 24603:2022(E)
Annex B (informative) Examples for methods for the establishment and culture of PSCs .22
Annex C (informative) Quality control procedure for biobanking of human and mouse PSCs .26
Bibliography .27
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ISO 24603:2022(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.
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ISO 24603:2022(E)
Introduction
Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells
(iPSCs), have been extensively studied in scientific research in order to improve the understanding
of developmental biology and diseases, to create organoids for drug screening, and to be applied in
cell-based therapies. In just a few years, thousands of PSC lines have been established in laboratories
around the world. PSC lines hold unique characteristics and behaviour due to their capability for both
self-renewal and differentiation into multiple cell types. However, the stem cell phenotype can be
changed by suboptimal cell culture technique, prolonged passage or changing the culture conditions.
Clearly the consequences of using adversely affected cells would be wasted time and resources but,
even more seriously, the generation of erroneous data in the literature which could both confuse
and delay scientific progress in this area. Accordingly, mouse PSCs have been used to establish our
fundamental understanding of PSC biology and these discoveries have been translated into human PSC
research to drive the development of new human-cell-based in vitro assays and potential regenerative
medicines. Mouse PSCs and human PSCs have become the most widely studied species in this field and
many significant scientific advances have been made by using PSCs from these two species. Of course,
PSC lines have been established from other species such as rat, porcine, canine, bovine, primate, etc.
and those from primates in particular have provided understanding of the biology of these cells which
can be more relevant to human stem cell biology than data from mouse PSCs. However, PSCs from
these species are much less used in research laboratories than mouse and human and are therefore not
described specifically in this document although much of this document will be relevant to them.
Human PSCs developed in research environments will give the clues to the development of cell therapies,
thus ensuring that cell lines used in this dynamic field have been prepared and documented appropriately
and have the correct identity and characteristics, which is critical to help ensure reproducibility in PSC-
based research. This document aims to meet the current demand for standardized PSC procedures
[9]
of biobanks and builds on international consensus agreed by PSC resource centres . This document
specifies the establishment, maintenance, characterization, storage and distribution requirements for
mouse and human PSCs, providing a general guideline for both biobanking and fundamental research
of PSCs.
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INTERNATIONAL STANDARD ISO 24603:2022(E)
Biotechnology — Biobanking — Requirements for human
and mouse pluripotent stem cells
1 Scope
This document specifies requirements for the biobanking of human and mouse pluripotent stem
cells (PSCs), including the collection of biological source material and associated data, establishment,
expansion, characterization, quality control (QC), maintenance, preservation, storage, thawing,
disposal, distribution and transport.
This document is applicable to all organizations performing biobanking with human and mouse PSCs
used for research and development.
This document does not apply to cell lines used for in vivo application in humans, clinical applications or
therapeutic use.
NOTE International, national or regional regulations or requirements, or multiple of them, 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 8601-1, Date and time — Representations for information interchange — Part 1: Basic rules
ISO 20387:2018, Biotechnology — Biobanking — General requirements for biobanking
ISO/TS 20388:2021, Biotechnology — Biobanking — Requirements for animal biological material
ISO 21709:2020, Biotechnology — Biobanking — Process and quality requirements for establishment,
maintenance and characterization of mammalian cell lines
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 20387:2018 and the following
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 https:// www .electropedia .org/
3.1
biobank
legal entity or part of a legal entity that performs biobanking (3.2)
[SOURCE: ISO 20387:2018, 3.5]
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ISO 24603:2022(E)
3.2
biobanking
process of acquisitioning and storing, together with some or all of the activities related to collection,
preparation, preservation, testing, analysing and distributing defined biological material as well as
related information and data
[SOURCE: ISO 20387:2018, 3.6]
3.3
cell line master file
complete dossier of all procedures and records used to generate and maintain a cell line
3.4
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.
[SOURCE: ISO 21709:2020, 3.3]
3.5
cell population purity
percentage of a particular cell type in a population, of which has the same specific biological
characteristics, such as cell surface markers, genetic polymorphisms and biological activities
[SOURCE: ISO/TS 22859:2022, 3.8]
3.6
cryopreservation
process by which cells are maintained in an ultra-low temperature in an inactive state so that they can
be revived later
[SOURCE: ISO 21709:2020/Amd 1:2021, 3.6]
3.7
differentiation
process to bring the cells into a defined cell state or fate
[SOURCE: ISO/TS 22859:2022, 3.11]
3.8
differentiation potential
ability that refers to the concept that stem and progenitor cells can produce daughter cells which are
able to further differentiate into other cell types
[SOURCE: ISO/TS 22859:2022, 3.12]
3.9
embryonic stem cell
ESC
pluripotent stem cell (3.21) derived from the inner cell mass of a blastocyst, i.e. an early stage pre-
implantation embryo
3.10
ethics review committee
body which is responsible for the evaluation and review of the ethical issues involved in the research
3.11
expansion
cell culturing process by which the cell number increases in vitro
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ISO 24603:2022(E)
3.12
feeder cell
mitotically inactivated cell used to support the growth of pluripotent stem cells (3.21)
3.13
genetic integrity
genome of cells that has not been altered
3.14
genetic state
phenotype of genetic profile of individual organism, including but not limited to karyotype (3.18),
integrity, mutation and knock-in of exogenous sequence
3.15
harvest
process of obtaining cells from a cell culture environment
3.16
identity verification
part of the process of verifying authenticity of a cell line in which cell origin is genetically confirmed
[SOURCE: ISO 21709:2020, 3.10]
3.17
induced pluripotent stem cell
iPSC
pluripotent stem cell (3.21) that is generated from somatic cells through artificial reprogramming by
the introduction of genes or proteins, or via chemical or drug treatment
3.18
karyotype
characteristics of the chromosomes of a cell, including its number, type, shape and structure, etc.
3.19
passage
subculture
process of further culturing of cells in a culture vessel to provide higher surface area/volume for the
cells to grow
Note 1 to entry: A passage can be performed by harvesting an aliquot from the parent vessel and reseeding it into
another vessel.
[SOURCE: ISO/TS 22859:2022, 3.18]
3.20
passage number
number of subculturing that occurred
Note 1 to entry: For this document, P is understood as the starting population of the cells.
0
[SOURCE: ISO 21709:2020, 3.13, modified — Note 1 to entry added.]
3.21
pluripotent stem cell
PSC
stem cell (3.26) that can differentiate into all cell types of the body and is able to self-renew indefinitely
in vitro
Note 1 to entry: PSCs include embryonic stem cells (ESCs) (3.9) (including fertilization derived ESCs, somatic cell
nuclear-transferred stem cells (3.25), etc.) and induced pluripotent stem cell (iPSCs) (3.17).
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ISO 24603:2022(E)
Note 2 to entry: ESC-like cells can also be isolated by parthenogenetic division of oocytes or other haploid cell
sources, and these cells have many of the characteristics of ESCs. However, certain features of these pluripotent
cell types can require specific characterization approaches.
3.22
population doubling time
PDT
doubling time
time taken for cultured cell count to double
Note 1 to entry: The time is measured in hours.
[SOURCE: ISO 21709:2020, 3.8, modified — “population doubling time” and “PDT” added as the preferred
term. Note 1 to entry added.]
3.23
self-renewal
ability of stem cells (3.26) to divide symmetrically, forming two identical daughter stem cells
Note 1 to entry: Adult stem cells can also divide asymmetrically to form one daughter cell which can proceed
irreversibly to a differentiated cell lineage and ultimately lead to specialized functional differentiated cells, while
the other daughter cell still retains the characteristics of the parental stem cell.
[SOURCE: ISO/TS 22859:2022, 3.23]
3.24
separation
process of obtaining target cells from biological samples
3.25
somatic cell nuclear-transferred stem cell
embryonic stem cells (3.9) derived from in vitro transfer of a donor cell nucleus into an enucleated oocyte
3.26
stem cell
non-specialized cells with the capacity for self-renewal (3.23) and differentiation potential (3.8), which
can differentiate into one or more different types of specialized cells
Note 1 to entry: Based on potency, stem cells can be divided into: totipotent stem cell (3.29), pluripotent stem cell
(3.21), multipotent stem cell, oligopotent stem cells, and unipotent stem cells (see Annex A).
[SOURCE: ISO/TS 22859:2022, 3.24, modified — Note 1 to entry replaced.]
3.27
stem cell marker
protein or gene specifically expressed in stem cells (3.26), usually used to isolate and identify stem cells
Note 1 to entry: Stem cell markers vary depending on stem cell type.
3.28
teratoma
tumour containing representative differentiated tissues and cells from the three germ layers
3.29
totipotent stem cell
stem cell (3.26) that can differentiate into an intact new organism including embryonal and extra
embryonal cells
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ISO 24603:2022(E)
3.30
viability
attribute of being alive (e.g., metabolically active, capable of reproducing, have intact cell membrane, or
have the capacity to resume these functions)
[SOURCE: ISO 21709:2020, 3.17, modified — “as defined based on the intended use” deleted.]
4 Abbreviated terms
bFGF basic fibroblast growth factor
EMRO embryo research oversight
ESC embryonic stem cell
HBV hepatitis B virus
HCMV human cytomegalovirus
HCV hepatitis C virus
HIV human immunodeficiency virus
HLA human leukocyte antigen
HTLV human T-lymphotropic virus
IFU instructions for use
iPSC induced pluripotent stem cell
KLF4 krueppel-like factor 4
KSR knockout serum replacement
mLIF mouse leukaemia inhibitory factor
MTA materials transfer agreement
OCT4 octamer-binding transcription factor 4
OriP origin of replication
PBMC peripheral blood mononuclear cell
PSC pluripotent stem cell
QC quality control
SOX2 SRY (sex determining region Y)-box 2
SSEA3 stage-specific embryonic antigen 3
SSEA4 stage-specific embryonic antigen 4
SSEA1 stage-specific embryonic antigen 1
STR short tandem repeat
SV40LT Simian virus 40 large T
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ISO 24603:2022(E)
TP treponema pallidum
5 General requirements
5.1 General
The biobank shall follow ISO 20387 and ISO 21709, in addition to this document. ISO/TR 22758 can be
used as additional reference for the implementation of ISO 20387. For mouse PSCs, ISO/TS 20388 shall
also be followed.
The biobank shall establish criteria and procedures for the isolation, establishment, expansion, storage,
thawing and transport of PSCs.
A data analysis procedure shall be established, documented, implemented, regularly reviewed and
updated.
The biobank shall use validated and/or verified methods and procedures for activities pertaining to
PSCs in accordance with ISO 20387:2018, 7.9.2 and 7.9.3, at all stages of the biological material life cycle
(as defined in ISO 20387:2018, 3.29).
According to the characteristics of PSCs, procedures, QC documents for collection, separation,
expansion, storage, transportation and testing, and data analysis shall be established, documented,
implemented, regularly reviewed and updated.
5.2 Legal and ethical requirements
ISO 20387:2018, 4.1.6, 4.3, 7.2.3.4, 7.3.2.4, A.7 a), and ISO 21709:2020, 4.2, shall be followed. For mouse
PSCs, ISO/TS 20388:2021, 4.2, shall also be followed.
The biobank shall collect relevant information on ethical requirements, implement and ‎regularly
update them, where relevant.
It is important to recognize that PSC lines are potentially not acceptable for use in research or
development or both in some countries, and shipment of cells to collaborating organizations will require
consideration of these differences. The biobank shall establish, document and implement policies on the
procurement and supply of PSCs.
Experimental plans using or establishing human PSCs should be consulted in a specialized ethics
review committee with particular expertise in topics relevant to the type and intended use of the PSC
lines in the biobank.
The biobank shall establish a process to verify and document cell line provenance, to be able to provide
evidence of ethical and regulatory compliance.
The biobank shall be aware whether reimbursement was made for the donation of human embryos/
tissues and whether the human embryo was created for research as this can be illegal in some countries.
For derivation of new pluripotent cell lines from human embryos, the ethical review process shall refer
to relevant expert ethical reviews.
[10]
EXAMPLE The human embryo research oversight (EMRO) process (ISSCR guidelines 2016, Chapter 2.1) .
Ethical requirements relevant for distribution are provided in 18.1.
5.3 Personnel, facilities and equipment
ISO 20387:2018, Clause 6, and ISO 21709:2020, 4.3, 4.4, 4.7, shall be followed. For mouse PSCs,
ISO/TS 20388:2021, 4.3, shall also be followed.
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ISO 24603:2022(E)
The biobank personnel shall be appropriately and specifically trained in PSC generation,
characterization, culture, cryopreservation, thawing and transport.
The biobank shall ensure that external operators providing PSC services demonstrate relevant
knowledge, experience and corresponding skills.
The biobank shall ensure that facilities, equipment and environmental conditions do not adversely
affect PSC quality attributes or invalidate the test results.
Equipment management procedures should be established, including the use of equipment and
maintenance plan.
The biobank shall control the operating environment and conditions (e.g. temperature, humidity,
cleanliness) according to the relevant characteristics of PSCs and the need for aseptic processing.
5.4 Reagents, consumables and other supplies
ISO 21709:2020, 4.5, shall be followed. For mouse PSCs, ISO/TS 20388:2021, 5.1.3, shall also be followed.
The biobank shall establish acceptance criteria for materials, including reagents and consumables,
necessary for PSC isolation, establishment, expansion, preservation, storage, thawing and transport.
If animal serum is used for PSC culture, there should be no evident potential high risk source of virus
or bovine spongiform encephalopathy, which cannot be managed by a risk assessment of the biological
source material and decontamination (such as irradiation for certain viruses).
For culture of human cell lines, if there are blood components in the culture medium (such as platelet
lysate, serum, albumin, transferrin and various cytokines), the source, batch number and quality
verification report shall be documented; and, if possible, a risk assessment shall be completed following
communication with the manufacturer on the risk of microbial contamination and other potential
hazards such as toxic contaminants. Where approved sources of these components are availab
...