Differentiation of porcine adipose‐derived stem cells into glucose responsive insulin‐secreting pancreatic islet‐like clusters

Abstract

While its ease of isolation, abundant source and multipotency have made the adipose‐derived stem cell (ADSC) a promising system for cell therapy and regenerative medicine, the approach on utilizing porcine ADSC to generate insulin‐secreting cells has not been explored. In our current study, subcutaneous adipose tissue on the back were dissected from 9‐day‐old male piglets (Y × L) purchased from a local pig farm and processed as previously described to isolate the stromal‐vascular fraction (SVF). Cells were then cultured in differentiation medium based on serum‐free DMEM/F12 as modified from Timper et al.. Our results showed that under light microscope, the morphology of pancreatic islet‐like clusters were observed at day 3 of culture, the function of insulin production identified by immunocytochemical analysis with confocal microscope and the expression of pancreatic β‐cell differentiation‐associated genes, including Pdx1, Pax4, Islet1, glucokinase, glucagon and Glut2, by RT‐PCR. Moreover, upon glucose challenge, the clusters secreted significantly higher amount of insulin as measured by ELISA on day 9 in a dose‐dependent manner compared to the basal level (5.5 mM glucose). Our present study, therefore, demonstrates the success of isolating and differentiating porcine ADSC into glucose responsive insulin‐secreting clusters. Their potentials in ameliorating type I diabetes in mouse and pig models are currently under investigation.