Skip to content
Education
Isle of NAM
GCCP

GCCP

Table of Contents

Definition

Good Cell Culture Practice (GCCP) is a guidance framework defining minimum quality standards for the handling, characterization, maintenance, and use of cells and tissues cultured in vitro. The framework was produced by the second ECVAM Task Force on Good Cell Culture Practice and published in 2005 as the founding GCCP guidance1; a substantially revised edition, GCCP 2.0, was published in 20222.

GCCP is not a biological model in itself; it is the quality system applied to every such model. Its biological basis lies in the intrinsic instability of living cells in vitro: over serial passage, cultured cells drift genetically and phenotypically, may dedifferentiate, transform, or senesce, can be cross-contaminated or misidentified, and remain susceptible to covert microbial infection, most notably by mycoplasmas. Each of these processes silently alters what a culture actually models1. GCCP therefore governs the cellular and structural mechanics that determine model identity: the characterization and maintenance of the essential morphological, functional, and genotypic characteristics of the culture system; the provenance and traceability of cell stocks; and the control of the media, sera, substrates, and culture parameters on which phenotype depends1 2.

Purpose

The primary value proposition of GCCP is that it maximizes the reproducibility, reliability, credibility, acceptance, and proper application of all results produced with in vitro systems1. It replaces a legacy situation in which cell culture practice was governed by local, unstandardized laboratory custom, and it was designed explicitly to complement, rather than replace, existing guidance and regulations such as Good Laboratory Practice1.

The framework became indispensable as in vitro systems expanded into regulatory safety testing and high-throughput screening1, and the 2022 revision was prompted by the diversification of cell models into induced pluripotent stem cell-derived models, three-dimensional cultures, organoids, and microphysiological systems2. The human relevance of GCCP is direct: it quality-assures the human cells and tissues on which human-relevant New Approach Methodologies depend, and its principles underpin the OECD guidance that reduces the uncertainties of cell and tissue-based predictions in chemical safety assessment3.

Methodology

GCCP 1.0 was constructed around six operational principles: characterization and maintenance of essential characteristics of cell culture systems; quality assurance; recording; safety; education and training; and ethics1. GCCP 2.0 restates these as six main principles — characterization and maintenance of essential characteristics, quality management, documentation and reporting, safety, education and training, and ethics — and extends their application to modern complex culture formats2.

Implementation relies on the following techniques and tools:

  • Characterization: authentication of cell line identity, notably by short tandem repeat profiling, together with verification of morphology, viability, and function, and confirmation of the absence of adventitious agents1 2.
  • Banking and passage control: controlled two-tier cell banking with master and working stocks, cryopreservation under documented conditions, and limits on passage number to contain genetic and phenotypic drift1 2.
  • Contamination control: routine aseptic technique and periodic mycoplasma screening by polymerase chain reaction, microbiological culture, or DNA staining, with elimination or disposal of infected cultures2.
  • Materials control: qualification and lot-to-lot testing of media, sera, substrates, extracellular matrices, growth factors, and reagents, with documented storage conditions and full traceability1 2.
  • Documentation: standard operating procedures for every manipulation, comprehensive records of provenance, passage history, and deviations, and archiving sufficient to permit reconstruction of the experiment1 2.
  • Training, safety, and ethics: documented competence of personnel, biosafety measures for operators and the environment, and ethical sourcing of human and animal material1 2.

For regulatory contexts, the OECD Good In Vitro Method Practices (GIVIMP) guidance operationalizes these principles into ten aspects, spanning roles and responsibilities, quality considerations, facilities, apparatus and reagents, test systems, test and control items, standard operating procedures, performance of the method, reporting of results, and storage and retention of records and materials3. For stem cells, characterization requirements extend to pluripotency markers, differentiation competency, and genomic stability monitoring4; organoids and microphysiological systems require additional batch quality control and functional benchmarking5.

Performance

The empirical case for GCCP rests on documented quality failures. More than 450 cell lines are registered as misidentified or cross-contaminated, and a conservative estimate indicates that in excess of 30,000 publications remain in circulation reporting data obtained with the wrong cells6. Survey data compiled by Drexler and Uphoff show mycoplasma contamination in a substantial proportion of laboratory cell lines, with reported institutional rates ranging from the low teens to seventy percent of cultures tested7. These persistent failure modes led the OECD to adopt GIVIMP explicitly as a tool to avoid a reproducibility crisis in in vitro toxicological science3.

With respect to reproducibility metrics, GCCP-aligned documentation and reporting are intended to permit inter-laboratory reconstruction of experiments; the dedicated reporting-standards initiative was launched because in vitro research still lacked generally accepted reporting criteria8.

Current technical boundaries are as follows. GCCP is guidance rather than binding regulation: adherence is voluntary, and adoption remains incomplete across academic and industrial laboratories2 8. GCCP 2.0 itself anticipates further revisions as science and technology evolve2. The legacy of misidentified and contaminated literature persists without correction or warning6, and reporting practice is not yet harmonized8.

Applications

  • Basic and biomedical research: GCCP constitutes the general quality baseline for all work with cells and tissues in vitro, from finite and continuous cell lines to primary cultures and stem-cell-derived systems1.
  • Regulatory toxicology and chemical safety assessment: GIVIMP applies GCCP principles to in vitro methods developed for regulatory use in human safety assessment and was adopted as OECD guidance in 20183; a second edition was published in December 20259.
  • Stem cell research, testing, and product development: the stem-cell expansion of GCCP provides quality assurance for the diverse cell types and culture formats of stem cell technology, from research through manufacture4.
  • Organoids and microphysiological systems: advanced GCCP addresses the quality issues specific to human primary, stem-cell-derived, and organoid models as well as microfluidic organ-on-chip systems5.
  • Scientific publishing and reporting: the GIVReSt initiative promotes complete and structured reporting of cell culture experiments; a draft guidance was published for stakeholder discussion in 20258 10.

Deployment stage and readiness: GCCP 1.0 (2005) and GCCP 2.0 (2022) are established and internationally recognized guidance documents, and GIVIMP is an adopted OECD guidance now in its second edition1 2 3 9. The framework is mature for conventional two-dimensional culture, while its application to organoids, microphysiological systems, and other complex models continues to evolve2 5; the reporting strand remains at the stakeholder consultation stage10.

Variations

  1. Coecke, S., Balls, M., Bowe, G., et al., Guidance on Good Cell Culture Practice. A report of the second ECVAM Task Force on Good Cell Culture Practice, Alternatives to Laboratory Animals 33(3), 261-287, 2005.
    The founding GCCP guidance document, produced by the second ECVAM Task Force with broad international participation, defines six operational principles covering the characterization and maintenance of essential characteristics of cell culture systems, quality assurance, recording, safety, education and training, and ethics. It sets minimum standards intended to maximize the reproducibility, reliability, credibility, and acceptance of all work with cells and tissues in vitro, and it was designed to complement, not replace, existing guidance and regulations. ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎

  2. Pamies, D., Leist, M., Coecke, S., et al., Guidance document on Good Cell and Tissue Culture Practice 2.0 (GCCP 2.0), ALTEX - Alternatives to Animal Experimentation 39(1), 30-70, published online 9 December 2021.
    GCCP 2.0 is the updated guidance document from GCCP 1.0, developed for practical use in the laboratory to assure the reproducibility of cell-based in vitro work. The update was essential because cell models have diversified into induced pluripotent stem cells, organoids, three-dimensional cultures, and microphysiological systems. The document describes six main principles — characterization and maintenance of essential characteristics, quality management, documentation and reporting, safety, education and training, and ethics — and explicitly foresees further revisions as science and technology evolve. ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎

  3. OECD, Guidance Document on Good In Vitro Method Practices (GIVIMP), OECD Series on Testing and Assessment No. 286, OECD Publishing, Paris, 10 December 2018.
    The OECD GIVIMP guidance applies good scientific, technical, and quality practices to the development and implementation of in vitro methods for regulatory use in human safety assessment. It addresses ten aspects of in vitro work, from roles and responsibilities through storage and retention of records and materials, and it was developed as a reference for best practices and as a tool to avoid a reproducibility crisis in in vitro toxicological science. ↩︎ ↩︎ ↩︎ ↩︎ ↩︎ ↩︎

  4. Pamies, D., Bal-Price, A., Simeonov, A., et al., Good Cell Culture Practice for stem cells and stem-cell-derived models, ALTEX - Alternatives to Animal Experimentation 34(1), 95-132, 2017.
    This document expands the original 2005 GCCP guidance to the increasingly diverse cell types and culture formats used in research, product development, testing, and manufacture, with particular attention to pluripotent stem cells and stem-cell-derived models. It specifies characterization requirements including pluripotency markers, differentiation competency, and genomic stability monitoring, and it addresses quality problems such as cross-contamination and microbial infection that compromise stem cell work. ↩︎ ↩︎ ↩︎

  5. Pamies, D., Bal-Price, A., Chesné, C., et al., Advanced Good Cell Culture Practice for human primary, stem cell-derived and organoid models as well as microphysiological systems, ALTEX - Alternatives to Animal Experimentation 35(3), 353-378, 2018.
    This guidance extends GCCP to the complex in vitro models that have emerged since 2005, including human primary cells, stem-cell-derived models, organoids, and microphysiological systems. It identifies the quality issues specific to these formats — batch variability, maturation state, functional heterogeneity, and the qualification of dynamic culture conditions — and defines characterization and control measures appropriate to each. ↩︎ ↩︎ ↩︎ ↩︎

  6. Horbach, S. P. J. M. and Halffman, W., The ghosts of HeLa: How cell line misidentification contaminates the scientific literature, PLOS ONE 12(10), e0186281, October 2017.
    This study estimates the scale of the literature contaminated by cell line misidentification. The authors report that more than 450 cell lines are known to be misidentified, frequently through cross-contamination with HeLa or other lines, and they conservatively estimate that in excess of 30,000 publications remain in circulation reporting results obtained with the wrong cells, without warning or correction. They argue for a fair notification system that cautions readers when interpreting this contaminated literature. ↩︎ ↩︎

  7. Drexler, H. G. and Uphoff, C. C., Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention, Cytotechnology 39, 75-90, 2002.
    This review compiles survey data on the incidence and sources of mycoplasma contamination in cell culture, its wide-ranging effects on eukaryotic cells, and the methods available for detection, elimination, and prevention. Reported contamination rates vary widely across institutions, from the low teens to seventy percent of cultures tested, which makes routine screening, together with robust elimination and prevention procedures, a core element of good culture practice. ↩︎

  8. Hartung, T., de Vries, R., Hoffmann, S., et al., Toward Good In Vitro Reporting Standards, ALTEX - Alternatives to Animal Experimentation 36(1), 3-17, 2019.
    This article summarizes the initiatives established to improve the quality of in vitro work and its reporting, observing that in vitro research still lacks generally accepted reporting criteria. It proposes Good In Vitro Reporting Standards (GIVReSt) so that cell culture experiments are reported completely enough to be reproduced, compared, and appraised by authors, reviewers, journals, and regulators alike. ↩︎ ↩︎ ↩︎ ↩︎ ↩︎

  9. OECD, Guidance Document on Good In Vitro Method Practices (GIVIMP), Second Edition, OECD Series on Testing and Assessment, OECD Publishing, Paris, 15 December 2025.
    The second edition of the GIVIMP guidance retains the ten aspects of good in vitro practice and updates the document with additional content, notably on ethical issues associated with the use of human serum. It reflects the continued evolution of quality expectations for in vitro methods used in regulatory safety assessment. ↩︎ ↩︎ ↩︎

  10. Mohapatra, R., Leist, M., von Aulock, S. and Hartung, T., Guidance for Good In Vitro Reporting Standards (GIVReSt) - A draft for stakeholder discussion and background documentation, ALTEX - Alternatives to Animal Experimentation 42(3), 376-396, 2025.
    This draft guidance operationalizes the GIVReSt initiative by proposing minimum reporting standards for cell culture experiments, covering monolayers, stem cells, organoids, and microphysiological systems. It builds on GCCP 2.0 and OECD reporting practice, and it addresses incomplete reporting of methodology as a major factor impairing reproducibility between laboratories. ↩︎ ↩︎ ↩︎