Serum and supplement optimization for EU GMP‐compliance in cardiospheres cell culture

Journal of Cellular and Molecular Medicine

Isotta Chimenti²  Roberto Gaetani¹  Elvira Forte¹  Francesco Angelini¹  Elena De Falco²  Giuseppe Biondi Zoccai²  Elisa Messina¹  Giacomo Frati² 

[1] Department of Molecular Medicine, Pasteur Institute – Cenci Bolognetti Foundation, “Sapienza” University of Rome, Rome, Italy;Department of Medical Surgical Sciences and Biotechnology, “Sapienza” University of Rome, Latina, Italy

关键词: adult stem cells;    cardiac cell therapy;    cardiospheres;    Good Manufacturing Practice compliance;    human sera;   

DOI  :  10.1111/jcmm.12210

来源: Wiley

【 摘 要 】

Abstract

Cardiac progenitor cells (CPCs) isolated as cardiospheres (CSs) and CS-derived cells (CDCs) are a promising tool for cardiac cell therapy in heart failure patients, having CDCs already been used in a phase I/II clinical trial. Culture standardization according to Good Manufacturing Practices (GMPs) is a mandatory step for clinical translation. One of the main issues raised is the use of xenogenic additives (e.g. FBS, foetal bovine serum) in cell culture media, which carries the risk of contamination with infectious viral/prion agents, and the possible induction of immunizing effects in the final recipient. In this study, B27 supplement and sera requirements to comply with European GMPs were investigated in CSs and CDCs cultures, in terms of process yield/efficiency and final cell product gene expression levels, as well as phenotype. B27− free CS cultures produced a significantly reduced yield and a 10-fold drop in c-kit expression levels versus B27+ media. Moreover, autologous human serum (aHS) and two different commercially available GMP AB HSs were compared with standard research-grade FBS. CPCs from all HSs explants had reduced growth rate, assumed a senescent-like morphology with time in culture, and/or displayed a significant shift towards the endothelial phenotype. Among three different GMP gamma-irradiated FBSs (giFBSs) tested, two provided unsatisfactory cell yields, while one performed optimally, in terms of CPCs yield/phenotype. In conclusion, the use of HSs for the isolation and expansion of CSs/CDCs has to be excluded because of altered proliferation and/or commitment, while media supplemented with B27 and the selected giFBS allows successful EU GMP-complying CPCs culture.

【 授权许可】

CC BY   
© 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.