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Quality Control Assays for Clinical-Grade Human Mesenchymal Stromal Cells: Methods for ATMP Release

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Mesenchymal Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1416))

Abstract

Mesenchymal stromal/stem cells (MSC) are promising candidates for the development of cell-based therapies for various diseases and are currently being evaluated in a number of clinical trials (Sharma et al., Transfusion 54:1418–1437, 2014; Ikebe and Suzuki, Biomed Res Int 2014:951512, 2014). MSC for therapeutic applications are classified as advanced therapy medicinal products (ATMP) (Regulation (EC) No 1394/2007 of the European Parliament and of the Council of 13 November 2007 on advanced therapy medicinal products and amending Directive 2001/83/EC and Regulation (EC) No 726/2004) and must be prepared according to good manufacturing practices (http://ec.europa.eu/health/documents/eudralex/vol-4). They may be derived from different starting materials (mainly bone marrow (BM), adipose tissue, or cord blood) and applied as fresh or cryopreserved products, in the autologous as well as an allogeneic context (Sharma et al., Transfusion 54:1418–1437, 2014; Ikebe and Suzuki, Biomed Res Int 2014:951512, 2014; Sensebé and Bourin, Transplantation 87(9 Suppl):S49–S53, 2009). In any case, they require an approved and well-defined panel of assays in order to be released for clinical use.

This chapter describes analytical methods implemented and performed in our cell factory as part of the release strategy for an ATMP consisting of frozen autologous BM-derived MSC. Such methods are designed to assess the safety (sterility, endotoxin, and mycoplasma assays) and identity/potency (cell count and viability, immunophenotype and clonogenic assay) of the final product. Some assays are also applied to the biological starting material (sterility) or carried out as in-process controls (sterility, cell count and viability, immunophenotype, clonogenic assay).

The validation strategy for each analytical method is described in the accompanying Chapter 20.

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References

  1. Sharma RR, Pollock K, Hubel A et al (2014) Mesenchymal stem or stromal cells: a review of clinical applications and manufacturing practices. Transfusion 54:1418–1437

    Article  CAS  PubMed  Google Scholar 

  2. Ikebe C, Suzuki K (2014) Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols. Biomed Res Int 2014:951512

    Article  PubMed  PubMed Central  Google Scholar 

  3. Sensebé L, Bourin P (2009) Mesenchymal stem cells for therapeutic purposes. Transplantation 87(9 Suppl):S49–S53

    Article  PubMed  Google Scholar 

  4. Regulation (EC) No 1394/2007 of the European Parliament and of the Council of 13 November 2007 on advanced therapy medicinal products and amending Directive 2001/83/EC and Regulation (EC) No 726/2004

    Google Scholar 

  5. EudraLex – volume 4 – good manufacturing practice (GMP) guidelines – part I – basic requirements for medicinal products. http://ec.europa.eu/health/documents/eudralex/vol-4

  6. Guideline on human cell-based medicinal products. EMEA/CHMP/410869/2006

    Google Scholar 

  7. Rayment EA, Williams DJ (2010) Concise review: mind the gap: challenges in characterizing and quantifying cell- and tissue-based therapies for clinical translation. Stem Cells 28:996–1004

    PubMed  PubMed Central  Google Scholar 

  8. EudraLex – volume 4 – good manufacturing practice (GMP) guidelines – annex 1 – manufacture of sterile medicinal products. http://ec.europa.eu/health/documents/eudralex/vol-4

  9. Gnecchi M, Melo LG (2009) Bone marrow-derived mesenchymal stem cells: isolation, expansion, characterization, viral transduction, and production of conditioned medium. Methods Mol Biol 482:281–294

    Article  CAS  PubMed  Google Scholar 

  10. Wagey R, Short B (2013) Isolation, enumeration, and expansion of human mesenchymal stem cells in culture. Methods Mol Biol 946:315–334

    Article  CAS  PubMed  Google Scholar 

  11. Roseti L, Serra M, Bassi A (2015) Standard operating procedure for the good manufacturing practice-compliant production of human bone marrow mesenchymal stem cells. Methods Mol Biol 1283:171–186

    Article  CAS  PubMed  Google Scholar 

  12. Guideline on potency testing of cell based immunotherapy medicinal products for the treatment of cancer, EMEA/CHMP/410869/2006

    Google Scholar 

  13. Wuchter P, Bieback K, Schrezenmeier H et al (2015) Standardization of good manufacturing practice-compliant production of bone marrow-derived human mesenchymal stromal cells for immunotherapeutic applications. Cytotherapy 17:128–139

    Article  CAS  PubMed  Google Scholar 

  14. Dominici M, Le Blanc K, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317

    Article  CAS  PubMed  Google Scholar 

  15. Commission Directive 2006/17/EC implementing Directive 2004/23/EC of the European Parliament and of the Council as regards certain technical requirements for the donation, procurement and testing of human tissues and cells

    Google Scholar 

  16. Directive 2004/23/EC of the European Parliament and of the Council on setting standards of quality and safety for the donation, procurement, testing, processing, preservation, storage and distribution of human tissues and cells

    Google Scholar 

  17. European Pharmacopoeia 8.0, Section 2.6.27 (Microbiological control for cellular products) (2014) European Directorate for the Quality of Medicines & HealthCare, Strasbourg, FR

    Google Scholar 

  18. European Pharmacopoeia 8.0, Section 2.6.1 (Sterility) (2014)

    Google Scholar 

  19. European Pharmacopoeia 8.0, Section 2.6.14 (Bacterial endotoxins) (2014) European Directorate for the Quality of Medicines & HealthCare, Strasbourg, FR

    Google Scholar 

  20. Endosafe – PTS Product Validation (2007) Endosafetimes – volume 13, n. 1 – Charles River Laboratories

    Google Scholar 

  21. European Pharmacopoeia 8.0, Section 2.6.7 (Mycoplasmas) (2014) European Directorate for the Quality of Medicines &HealthCare, Strasbourg, FR

    Google Scholar 

  22. European Pharmacopoeia 8.0, Section 2.7.24 (Flow Cytometry) (2010) European Directorate for the Quality of Medicines &HealthCare, Strasbourg, FR

    Google Scholar 

  23. US Department of Health (2008), Guidance for FDA reviewers and sponsors: content and review of chemistry, manufacturing, and control (CMC) information for human gene therapy investigational new drug applications (INDs). US Department of Health, FDA, CBER

    Google Scholar 

  24. Castro-Malaspina H, Ebell W, Wang S (1984) Human bone marrow fibroblast colony-forming units (CFU-F). Prog Clin Biol Res 154:209–236

    CAS  PubMed  Google Scholar 

  25. Strober W (2001) Trypan blue exclusion test of cell viability. Curr Protoc Immunol Appendix 3B

    Google Scholar 

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Acknowledgments

This work was supported by the Fondazione Cardiocentro Ticino—Lugano. The authors would like to acknowledge Maura Filippini for her excellent editing assistance.

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Authors and Affiliations

  1. Lugano Cell Factory, Cardiocentro Ticino - Swiss Institute of Regenerative Medicine (SIRM), Via Tesserete 48, Lugano, 6900, Switzerland

    Marina Radrizzani, Sabrina Soncin, Viviana Lo Cicero, Gabriella Andriolo, Sara Bolis & Lucia Turchetto

Authors
  1. Marina Radrizzani
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  2. Sabrina Soncin
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  3. Viviana Lo Cicero
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  4. Gabriella Andriolo
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  5. Sara Bolis
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  6. Lucia Turchetto
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Corresponding author

Correspondence to Lucia Turchetto .

Editor information

Editors and Affiliations

  1. Dept of Molecular Med,Unit of Cardiology, University of Pavia, PAVIA, Italy

    Massimiliano Gnecchi

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© 2016 Springer Science+Business Media New York

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Radrizzani, M., Soncin, S., Lo Cicero, V., Andriolo, G., Bolis, S., Turchetto, L. (2016). Quality Control Assays for Clinical-Grade Human Mesenchymal Stromal Cells: Methods for ATMP Release. In: Gnecchi, M. (eds) Mesenchymal Stem Cells. Methods in Molecular Biology, vol 1416. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3584-0_19

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  • DOI: https://doi.org/10.1007/978-1-4939-3584-0_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3582-6

  • Online ISBN: 978-1-4939-3584-0

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