Preparation Of Adipose-derived Stem Cells (ASCs) In Various Sources Of Human Ascs And Its Directional Differentiation Into Pancreatic ? Cells

Objective:Adipose-tissue derived Stem Cells(ASCs)are one of the main stem cell sources in regeneration medicine because of their stemness,capacity of inducible differentiation and the potential for therapeutic effect.Here,we researched the alternative sources of adipose tissues and established the isolation and culture conditions for ASCs.In particularly,we collected abundant number of adipose tissues from Mensenteric and Visceral during bariatric surgery,and established the method of isolation and culture of ASCs derived from three depots of the body.Furthermore,we explore the chemical way of ASCs differentiate to ? cells to apply to the clinic more safely.Methods:We isolated the ASCs from different depots by different collagenase I concentration,digestion time etc.;we cultured the isolated ASCs with three different mediums;we tested the osteogenesis,adipogenesis and chondrogenic differentiation;we detected the purity by FACS.By q PCR,we detected the cell type markers in FACS collected cells.We directed the differentiated of ASCs to pancreatic ?-cells through chemical induction and detected the key transcriptional factors expression during the differentiation process.Results:1,Different isolation methods and culture conditions were applied for various depots of ASCs.The culture condition 1 is the best for all sources of ACSs.2,ASCs derived from subcutaneous show the best differentiation ability.3,ASCs in subcutaneous source show the high purity.Visceral ASCs isolated from nearby of omentum were mixed with epithelial cells and vascular endothelial cells.4,Tested once the current protocol,for induction of ASCs into pancreatic beta cells,the expression of beta cell specific genes can be detected during the end differentiation stages.Conclusion:For ASCs isolated from various sources,culture condition 1 is the best.subcutaneous ASCs have the best differentiation ability,and show highest purity.Differentiation of ASCs to pancreatic beta cells needs further exploration and optimization to achieve optimal efficiency.