Study On Treatment Effects Of Human Placenta-derived Stem Cells And Its Islet-like Cells Induced In Vitro On Gestational Diabetic Rats

purposeIn this study,human placenta-derived mesenchymal stem cells(hPDMSCs)were induced into islet-like cells(ILCs)in vitro,and two types of cells were respectively implanted into gestational diabetes mellitus(GDM)rats.The levels of blood glucose,adiponectin,tumor necrosis factor a(TNF-a)and interleukin-6(IL-6)in GDM rats were detected to investigate the treatment effects of hPDMSCs and ILCs on gestational diabetes mellitus,providing a theoretical basis for the future treatment of gestational diabetes mellitus by stem cells.methodMesenchymal stem cells(MSCs)were isolated from placenta of healthy full-term cesarean section women,then isolated,cultured and identified.The identified MSCs were induced into islet-like cells in vitro.The induced islet-like cells were stained with Dithizone(DTZ)to determine the specificity of islet cells.50 female SD rats and 25 male SD rats were selected.10 female rats were fed with normal diet and another 40 female rats were fed with high sugar and fat.Four weeks later,male and female rats mated at a ratio of 1:2.Forty female rats were injected with streptozotocin(STZ)after conception.Fasting blood sugar>16.7 mmol/L was the successful model of GDM rats.10 normal-fed female rats were divided into normal pregnant rats group(group A).The remaining 40 GDM rats were randomly divided into four groups:non-intervention of GDM rats group(group B),physiological saline intervention of GDM rats group(group C),human placenta-derived stem cells intervention(non-induction)of GDM rats group(group D),and islet-like cells intervention of GDM rats group(group E).Fasting glucose was measured on the 6th,12th,and 18th gestation,and serum adiponectin,TNF-a,and IL-6 were measured on the day of cesarean section(the 18th gestation).resultStem cells were extracted from placenta for passage and culture,and the third generation of hPDMSCs were identified by flow cytometry.The results showed that CD44 and CD 105 were positive.It was confirmed that the obtained cells were human placenta-derived mesenchymal stem cells and could be used for subsequent research.After the induction of hPDMSCs,with the extension of the induction time,the cell morphology changed from spindle shape to round and gathered to the center,and the stem cells after the induction differentiation for 28 days turned brown-red by dithizone staining,which proved that the induced cells were islet like cells.3.The GDM mouse model was established for female rats with fasting blood glucose>16.7mmol/L.The experiment was divided into five groups:normal pregnant rats group(group A),non-intervention of GDM rats group(group B),physiological saline intervention of GDM rats group(group C),human placenta-derived stem cells intervention(non-induction)of GDM rats group(group D),and islet-like cells intervention of GDM rats group(group E).Fasting blood glucose of all experimental groups was measured on the 6th,12th and 18th day of gestation.The blood glucose level of GDM rats in the 4 groups at the same time point was higher than that of the normal pregnant rats(group A),the difference was statistically significant(P<0.05).Blood glucose levels of the non-intervention group(group B)and physiologicall saline group(group C)showed no statistical significance(P>0.05).Compared with group B and C,the blood glucose level of hPDMSCs group(group D)and ILCs group(group E)decreased significantly(P<0.05).Blood glucose level of group E was lower than that of group D on the 12th and 18th day of gestation(P<0.05).Compared with that of group D and group E,blood glucose levels on the 12th day of gestation fell more than that of the 6th day,which presented statistical significance(P<0.05).However,there was no statistical difference between blood glucose levels on the 18th and 12th day of gestation(P>0.05).Compared with group A,the levels of adiponectin in group B and group C decreased,while the levels of TNF-α and IL-6 increased(P<0.05).The differences of adiponectin,TNF-a and IL-6 between group B and C showed no statistical significance(P>0.05).Compared with group B and C,the levels of adiponectin and TNF-a and IL-6 decreased in group D and E,with statistically significant difference(P<0.05).Compared with group D and E,group E had higher adiponectin level,lower TN F-a and IL-6 level,and the difference was statistically significant(P<0.05).conclusionBoth human placenta-derived stem cells and islet-like cells can reduce the blood glucose of GDM rats,reduce the levels of TNF-a and IL-6,and increase the level of adiponectin,which is effective in the treatment of GDM rats.2.Compared with human placenta-derived stem cells and islet-cells,islet-cells decreased blood glucose,TNF-a,IL-6 and increased adiponectin levels more significantly,and the effect of islet-cells on GDM rats was better than that of human placenta-derived stem cells.