Insulin Secretion Of Human Umbilical Cord Mesenchymal Stem Cells Transfected With A Mutative Adenoviral Human Proinsulin Gene

Background:At present,diabetes already has become being the third big death diseases only inferior to the heart blood vessel of brain illness and cancer.It is a disease affecting insulin and sugar content,divided into two main species types:type 1 and type 2.Their common characteristic is the pancreatic beta cells loss or the body can not produce enough insulin or effectively use insulin.Type 1 diabetes is an insulin-dependent,autoimmune disorder in which the insulin producing beta cells of the pancreatic islets are selectively destroyed.Islet transplant is considered as the optimal treatment for typeâ… diabetes.However,because of the need to provide a large number of donor islet sufficient to achieve the purpose of clinical diabetes,after transplantation at the same time,also has the immune rejection problems,these to some extent have hindered its development.Objective:Our study was based on the characteristics of diabetes,using human umbilical cord-derived mesenchymal stem cell characteristics,mutative human proinsulin gene transfected into human umbilical cord-derived mesenchymal stem cells,caused it to express the mature human insulin. Methods and Results:A novel approach to produce insulin-secreting cell products using human umbilical cord-derived mesenchymal stem cells(UC-MSCs) carrying a human proinsulin gene was developed in the present study.In this study,we first constructed an adenovirus vector with a mutated human proinsulin gene and then transfected UC-MSCs with the vector.The transfected UC-MSCs were analyzed for their biological characteristics including the immune phenotype,the ability of differentiation and the expression of furin protease in comparison with normal UC-MSCs.Transcription and expression of the proinsulin gene in transfected UC-MSCs were detected by RT-PCR,western blot and immunofluorescence.And the secretion of mature insulin and C-peptide was examined by ELISA.Conclusion:We generated the human proinsulin gene containing the recognition site of the furin protease,and then constructed it to the adenovirus vector system(pAdINS-M2).After pAdINS-M2 infected the UC-MSCs,human mature insulin and C-peptide were express.It may be provide some powerful experiment basis for the treatment of typeâ… diabetes.