Good manufacturing practice-compliant isolation and culture of human adipose-derived stem cells

Abstract

Adipose-derived stem cells (ADSCs) are excellent for regenerative medicine. Like mesenchymal stem cells, ADSCs possess multi-potent differentiation capacity that enables them to differentiate into osteoblasts, chondrocytes and adipocytes, as well as trans-differentiation into other cells. ADSC transplantation has gained attention in recent years, especially in vitro expanded ADSC transplantation. This study aimed to provide a new method to in vitro primarily culture and secondary culture of ADSCs that were compliant with good manufacturing practice for clinical applications. Stromal vascular fraction (SVF) was extracted from adipose tissue by commercial kits. SVF was expanded in vitro in medium with non-allogeneic supplements. Cultured ADSCs maintained immune-phenotype, karyotype, and differentiation potential after ten passages. Moreover, ADSCs at 15th passage could not form tumors in NOD/SCID mice. This research produced a suitable protocol for clinical applications of expanded ADSCs.