National foreword

This British Standard is the UK implementation of ISO 20012:2026.

The UK participation in its preparation was entrusted to Technical Committee BTI/1, Biotechnologies.

A list of organizations represented on this committee can be obtained on request to its committee manager.

Contractual and legal considerations

This publication has been prepared in good faith, however no representation, warranty, assurance or undertaking (express or implied) is or will be made, and no responsibility or liability is or will be accepted by BSI in relation to the adequacy, accuracy, completeness or reasonableness of this publication. All and any such responsibility and liability is expressly disclaimed to the full extent permitted by the law.

This publication is provided as is, and is to be used at the recipient’s own risk.

The recipient is advised to consider seeking professional guidance with respect to its use of this publication.

This publication is not intended to constitute a contract. Users are responsible for its correct application.

Compliance with a British Standard cannot confer immunity from legal obligations.

This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 March 2026.

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 document 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).

ISO draws attention to the possibility that the implementation of this document may involve the use of (a) patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a) patent(s) which may be required to implement this document. However, implementers are cautioned that this may not represent the latest information, which may be obtained from the patent database available at www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.

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.

Introduction

Natural killer (NK) cells, also known as large granular lymphocytes (LGL), are a type of cytotoxic lymphocytes belonging to the innate immune system. NK cells constitute 5 % to 15 % of the mononuclear cells and up to 20 % of lymphocytic population^[1^] in the peripheral blood. The expression patterns of activating receptors and inhibitory receptors determine the activating status and functionalities of NK cells. NK cells can eliminate pathogen-infected cells, cancerous cells, and other unhealthy cells by direct cytotoxicity. They express apoptosis-related ligands (TRAIL and FasL) and cytotoxic granules (granzymes and perforin) which can induce cell death in stressed cells. NK cells with the expression of CD16 can also kill antibody-coated target cells by antibody-dependent cell cytotoxicity (ADCC). As innate immune cells, NK cells are also engaged in reciprocal interactions with other immune cells to limit or exacerbate immune responses^[2^].

In adult, hematopoietic stem cells (HSCs) differentiate into common lymphoid progenitors (CLPs), then, give rise to NK progenitors (NKP). NKPs migrate into all lymphoid tissues or organs, and differentiate into mature NK cells subsequently^[3^]. However, NK cells can also arise from erythro-myeloid progenitors (EMPs) in the yolk sac. NK cells can be generated by induction from human pluripotent stem cells (hPSCs)^[4^]. Regarding the various tissue origins of natural NK cells during development, there also has been established various methods for generating induced NK cells from hPSCs. In the NK cell regeneration system, hPSCs are first induced into mesoderm progenitors or lateral plate mesoderm cells via embryo body (EB) formation or monolayer differentiation methods. Then, these cells can be induced into hemogenic endothelial cells (HECs) which further differentiate into hematopoietic progenitor cells (HPCs) via hematopoietic transition (EHT). NK cells can even totally regenerated from these HPCs under specified cytokine combinations^[5^],^[6^].

NK cells are fragile and sensitive to cryopreservation and thawing. Standardized approaches to maintain the functionality of banked NK cells is needed. hPSC-dervied NK cell are primary cells, not immortalized, and therefore have a finite life span.

This document is applicable for academic centers, public and private institutions performing NK cell generation from hPSCs (Research and Development) and preclinical studies, not for clinical use.

Importantly, this document is focused on NK cell regeneration that have been reported from hPSCs in culture for research purposes.

EXAMPLE Applications of NK cells (e.g. immunoresponse regulation, anti-tumor or anti-viral)

1 Scope

This document specifies requirements for the biobanking of human natural killer (NK) cells derived from human pluripotent stem cells (hPSCs), including the requirements for the differentiaton, culture, characterization, quality control, storage, thawing and transport of NK cells.

Requirements for the collection of biological source material, the transport to and reception of biological source material and hPSCs at the biobank, as well as the establishment, expansion and QC of hPSCs are covered in ISO 24603.

This document is applicable to all organizations performing biobanking of human NK cells used for research and development in the life sciences.

This document does not apply to human NK cells for the purpose of 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

3 Terms and definitions

4 Abbreviations

5 General requirements

5.1 General

5.2 Legal and ethical requirements

5.3 Personnel, facilities and equipment

5.4 Reagents, consumables and other supplies

5.5 Management of information and data

6 hPSCs and associated data

7 Generation of hPSC-NK cells from hPSCs

7.1 Processes

7.2 Unique identification

7.3 Testing for infectious agents

7.4 Generation of hPSC-NK cells and culture

7.5 Subculture and limited expansion

8 Characterization of hPSC-NK cells

8.1 General

8.2 Viability

8.3 Morphology

8.4 Population doubling time (PDT) and subculture/expansion

8.5 Cell population purity

8.6 Immunophenotyping by flow cytometry

8.7 Microbiological contamination

8.8 hPSC-NK cell activity and function

9 Quality control

10 Storage

10.1

10.2

10.3

10.4

10.5

10.6

10.7

11 Thawing and culturing

12 Disposal

13 Distribution of hPSC-NK cells — Information for users

14 Transport of hPSC-NK cells

14.1 General

14.2 Transport of frozen hPSC-NK cells

14.3 Transport of living hPSC-NK cells

Bibliography

BS ISO 20012:2026

Biotechnology. Biobanking. Requirements for human natural killer cells derived from pluripotent stem cells

Current

Published:

31 Mar 2026