Differentiation of Rat Adipose-Derived Mesenchymal Stromal Cells toward Insulin-Producing Cells

Background: Until recently, much effort has been directed towardfinding novel stem cell sources and improving differentiationprotocols to develop a promising therapeutic method fortreating T۱DM (Type ۱ Diabetes Mellitus). In this study, wepropose a novel three stage differentiation method for efficientdifferentiation of ATDMSCs to functional IPCs.Materials and Methods: ATDMSCs were isolated from ratadipose tissue using collagenase type ۱. ATDMSCs were differentiatedinto IPCs via a ۱۴-day basic protocol using ۱% insulintransferrin selenium (ITS) and ۱% nicotinamide in Dulbecco’sModified Eagle’s Medium. Insulin granule formation, glucosestimulatedinsulin secretion and gene expression pattern relatedto the pancreatic endocrine development and function were analyzedin obtained IPCs (Insulin Producing Cells). The diabeticrats (n=۱۰) were divided into two groups. One group (n=۵) receivedundifferentiated ADMSCs; another group (n=۵) was injectedwith differentiated IPCs. Then, ۱×۱۰۶ of the isolated cellswere injected to the tail vein of the rats. Fasting blood glucoseconcentrations were measured once a week. At the end of thesixth week after transplantation, the blood insulin concentrationswere determined.Results: The obtained IPCs exhibited significant expressionof MafA, Nkx۲.۲, Nkx۶.۱, Ngn۳, insulin, and Isl۱, and insulinsecretion ability in response to glucose challenge. Rats that receivedthe differentiated IPCs exhibited a higher ability to normalizeblood glucose and insulin secretion when compared tocontrol group.Conclusion: The functional IPCs were successfully obtainedfrom ATDMSCs using a three-step protocol. The potency ofthe differentiated IPCs for the secretion of insulin and glycemiccontrol showed a significant increase compared to the un-differentiatedATDMSCs in vivo. However the differentiated IPCswere unable to restoration of euglycemia and normal acute insulinresponse to glucose. Further examinations are requiredto improvement of the three stage differentiation protocol formore efficient cell therapy of diabetic patients in future.

کلیدواژه ها:

ATDMSCs ، IPCs ، T۱DM ، Stem Cells