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

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