Abstract
Effective harvesting procedure for adipose tissue is demanded by the affordable Good Manufacturing Practice-Compliant Production of clinical-grade adipose tissue-derived stem cells (hADSCs). Enzymatic digestion using collagenase is the most reliable method of adipose tissue-derived stem cells (hADSCs) isolation, while the optimized loading volume ratios of digestion to container during the shaking process of adipose tissue and collagenase mixture are still lacking. This study was conducted to determine the optimized loading volume ratio (mixture to container) for enzymatic digestion of Stromal/Stem Cells from lipoaspirate. Lipoaspirates were obtained from twelve women immediately after liposuction. Then tissue from each patient was divided into four groups according to different loading volume ratios in 50 ml centrifugal tube: 0.2 group, 0.4 group, 0.6 group, 0.8 group. Stromal vascular fractions (SVF) were obtained from each group, then total cell counts, viability and viable cell count were performed. hADSCs were harvested at passage (P) 2, whose morphologies, immunophenotypes, proliferation, and tri-differentiation abilities were compared. 0.4 loading volume ratio provided the highest cell yield, favorable viability and viable cell yield. The proliferation and triple differentiation ability of hADSCs obtained by 0.4 group was not inferior to that of other groups. Therefore, 0.4 may be the optimal loading volume ratio for hADSCs isolation from lipoaspirate by enzymatic digestion in current setting.