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. 2013 Jan 4;430(1):325-30.
doi: 10.1016/j.bbrc.2012.11.001. Epub 2012 Nov 9.

Good Manufacturing Practice-Compliant Animal-Free Expansion of Human Bone Marrow Derived Mesenchymal Stroma Cells in a Closed Hollow-Fiber-Based Bioreactor

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Good Manufacturing Practice-Compliant Animal-Free Expansion of Human Bone Marrow Derived Mesenchymal Stroma Cells in a Closed Hollow-Fiber-Based Bioreactor

Philipp Nold et al. Biochem Biophys Res Commun. .

Abstract

Mesenchymal stroma cells (MSC) are increasingly recognized for various applications of cell-based therapies such as regenerative medicine or immunomodulatory treatment strategies. Standardized large-scale expansions of MSC under good manufacturing practice (GMP)-compliant conditions avoiding animal derived components are mandatory for further evaluation of these novel therapeutic approaches in clinical trials. We applied a novel automated hollow fiber cell expansion system (CES) for in vitro expansion of human bone marrow derived MSC employing a GMP-compliant culture medium with human platelet lysate (HPL). Between 8 and 32 ml primary bone marrow aspirate were loaded into the hollow fiber CES and cultured for 15-27 days. 2-58 million MSC were harvested after primary culture. Further GMP-compliant cultivation of second passage MSC for 13 days led to further 10-20-fold enrichment. Viability, surface antigen expression, differentiation capacity and immunosuppressive function of MSC cultured in the hollow fiber CES were in line with standard criteria for MSC definition. We conclude that MSC can be enriched from primary bone marrow aspirate in a GMP-conform manner within a closed hollow fiber bioreactor and maintain their T lymphocyte inhibitory capacity. Standardized and reliable conditions for large scale MSC expansion pave the way for safe applications in humans in different therapeutic approaches.

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