Experimental Study Of Human Umbilical Source Islet Precursor Cell Transplantation For Type 1 Diabetes

Objective:To evaluate the effect of hUMSCs on T1 D rats after in vitro induction of pancreatic islet precursor cells.Methods: 1.hUMSCs were isolated,cultivated,and identified;2.The hUMSCs at passage 4 was induced by phased induction and the morphological changes of the cells were observed;3.Collected the cells on the 7th,14 th and 21 st day of the induction.Immunofluorescence was used to detect the expression of Ngn3?insulin and MafA;Western-blot was used to detect the expression of Glut2 and MafA;4.Forty-four healthy male SD rats were divided into four groups by simple random sampling: induced type cell group,stem cell type roup,model group and normal group.Except for the normal group,the rest were given STZ by intraperitoneal injection to establish T1 D model,the normal group were given physiological saline;5.The islet progenitor cells were labeled with CM-DIL and transplanted into the pancreas capsule of the diabetic rats.The experimental period was 60 days,the whole experiment is divided into 2W,4W,6W,8W four time periods;the changes of blood glucose and body weight were observed;HE dyeing was used to observe the islet morphology;The expression of MafA in each period of each group was detected by Western-blot;Analysis of MafA and blood glucose correlations after transplantation;Analysis of the difference in the expression of MafA between stem cell group and induced cell group.Results: 1.The hUMSCs were successfully isolated and cultured.After induced,cell morphology became broad and colony appeared;2.Immunofluorescence showed positive expression of Ngn3,insulin and MafA.Western-blot was used to detect the expression of MafA and Glut2,they increased gradually in the process of the induction,MafA in the 7d was almost not expressed,14 d and 21 d slightly increased;Glut2 increased at 7d,peaked at 14 d,slightly lower at 21 days;3.38 rats were successfully modeled.The blood glucose ofthe experimental group was increased on the 7th day.After the transplantation,the blood glucose of stem cells and induced cells decreased in different degrees,but the induction of cell group decreased more significantly,model group remained in hyperglycemia;The weight of the model group continued to decrease during the whole experiment,the weight of Induced cell group and stem cell group increased after 4 weeks;4.Western-blot showed that MafA expression was detected after transplantation and showed an increasing trend,the stem cell group was significantly elevated at 6W,the induction of cell group was significantly increased at 4W;There was a negative correlation between MafA and blood glucose correlation after transplantation;There was no significant difference in the expression of MafA between stem cell group and induced cell group(P> 0.05).Conclusion:hUMSCs can induce differentiation into islet progenitor cells in the induced microenvironment,the induced cells can reduce hyperglycemia and rise weight in the diabetic rats after transplantation.