Abstract
Background aims. The discovery of unrestricted somatic stem cells (USSC), a non-hematopoietic stem cell population, brought cord blood (CB) to the attention of regenerative medicine for defining more protocols for non-hematopoietic indications. We demonstrate that a reliable and reproducible method for good manufacturing practice (GMP)-conforming generation of USSC is possible that fulfils safety requirements as well as criteria for clinical applications, such as adherence of strict regulations on cell isolation and expansion. Methods. In order to maintain GMP conformity, the automated cell processing system Sepax (Biosafe) was implemented for mononucleated cell (MNC) separation from fresh CB. After USSC generation, clinical-scale expansion was achieved by multi-layered CellSTACKs (Costar/Corning). Infectious disease markers, pyrogen and endotoxin levels, immunophenotype, potency, genetic stability and sterility of the cell product were evaluated. Results. The MNC isolation and cell cultivation methods used led to safe and reproducible GMP-conforming USSC production while maintaining somatic stem cell character. Conclusions. Together with implemented in-process controls guaranteeing contamination-free products with adult stem cell character, USSC produced as suggested here may serve as a universal allogeneic stem cell source for future cell treatment and clinical settings.
Acknowledgments
We gratefully acknowledge the Deutsche Forschungsgemeinschaft (DFG) for funding the research group FOR 717, including the project Ko2119/6-1, and the German José Carreras Leukemia Foundation for supporting us with grant DJCLS-R07/05v. We would like to thank A. Lefort and D. Stapelkamp for their technical support.
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.