| ObjectiveIn this study,human placenta-derived mesenchymal stem cells(hPDMSCs)were induced to differentiate into islet-like cells(ILCs)in vitro.Then they were implanted into the gestational diabetes mellitus(GDM)pregnant mice through the tail vein.The blood glucose and serum insulin were used to determine the induction.The function of the living cells provides basic theoretical research basis for the clinical application of human placenta-derived stem cells in GDM patients.In addition,this experiment examined the possibility of human placenta-derived stem cells inducing differentiation into pancreatic islet-like cells to investigate the mechanism of action of human placenta-derived stem cells on gestational diabetes in anticipation of the diagnosis and treatment of GDM and postpartum type ? diabetes.Play a guiding role.MethodHuman placental tissue was obtained under aseptic conditions,and human placental derived mesenchymal stem cells were extracted therefrom for isolation and culture.Then,flow cytometric techniques were used to identify whether the isolated cultured cells expressed CD44 and CD 105,etc.,to determine that the isolated cells were mesenchymal stem cells.The identified stem cells were induced to differentiate into islet-like cells by containing a high-glucose medium and a low-glucose medium.The entire induction period required about 28 days.After induction of differentiation,cells were stained with dithizone(DTZ),and the specificity of islet cells was determined by diphenylthiocarbazide(DTZ)staining.40 female SD rats and 20 male SD rats were selected.They were divided into 10 normal pregnant rats,10 rats treated with GDM saline,10 rats without GDM,and 10 rats treated with GDC.Four weeks after the female rats in the GDM group were fed a high-fat and high-sugar diet,the rats were mated in a 2:1 ratio.Where there is visible sperm or a vaginal plug,it indicates that it has been pregnant.After two intraperitoneal injections of STZ(streptozotocin),two days apart.On the 4th day of pregnancy,fasting blood glucose was measured,and fasting blood glucose>16.7 mmol/L was established as a GDM rat model.To determine the success of the model,ILC injection.Fasting blood glucose was measured with a blood glucose meter two days after each transplant and before caesarean section(caesarean section on the 18th day of gestation).Changes in insulin levels were measured using the Elisa method.The morphological changes of the islet cells were observed with an ordinary microscope.Result1.By flow cytometry,third generation human placental mesenchymal stem cells positively express CD44,CD 105.The obtained third generation human placental mesenchymal stem cells can be used for stem cell intervention experiments.2.After 7 days of induction,the original dispersed cells began to aggregate.As the time went by,the number and size of cell clusters increased significantly.After induction for 28 days,dithizone was used to induce islet-like cells.Staining was performed to identify that the sarcomatous cells were stained red-brown,demonstrating that the cells were ILC.3.After the rats were modeled,fasting blood glucose was measured on the 4th day of gestation.There were 30 pregnant rats whose fasting blood glucose was>16.7 mmol/L.The GDM mouse model was established and was included in the experimental group,divided into four groups of ABCD.10 IGM intervention group of GDM rats(group A),10 untreated GDM rats(group B),10 GDM rats normal saline intervention group(group C),and 10 normal pregnant rats(group D).4.The blood glucose levels in GDM groups were higher than those in normal pregnant mice on days 6,12 and 18(P<0.01).GMC group(group A)was significantly decreased compared with group B and C.The difference was statistically significant(P<0.01).In group A,fasting blood glucose decreased progressively with the increase of the number of ILC implantation during the pregnancy;the blood glucose level of group A was lower than that of group B and C at the same time points,and the difference between groups was statistically significant(P<0.01).Blood glucose in group B and C did not change significantly during pregnancy.There was no significant difference between groups(P>0.05).The insulin levels in GDM groups at 6d,12d,and 18d were all lower than those in normal pregnant mice,and the difference was statistically significant(P<0.01).GDM group without intervention of ILC(B,C group)decreased more significantly than group A,the difference was statistically significant(P<0.01).In group A,during the pregnancy,with the increase of the number of ILC implantation,insulin gradually increased;in groups A and B,C,insulin levels increased at the same time point,and the difference between groups was statistically significant(P<0.01).There was no significant change in group B and C during pregnancy.There was no significant difference between groups(P>0.05).5.The number of pancreatic islets in each group of GDM was significantly reduced,the volume was atrophic,the morphology was irregular,the structure was loose,the exocrine cells and acini were destroyed,the boundary of the cells was unclear,a large number of lymphocytes infiltrated,and the number of islet cells decreased under high magnification.Impaired pancreatic tissue cells in the ILC intervention group(group A)were repaired,pancreatic tissue atrophy was reversed,vacuoles were reduced,and inflammatory infiltrates were reduced.Conclusion1.The experiment succeeded in extracting placental mesenchymal stem cells from human placenta tissue and observing the growth and fusion of the placenta through a microscope.It was confirmed that it has the characteristics of mesenchymal stem cells and can be isolated,cultured,and expressed in vitro.Surface-related antibodies.2.In vitro,hPDMSCs were induced to differentiate into islet-like cells and stained with a dithizone stain to a red-brown color,confirming that the induced cells were islet-like cells.3.Islet-like cells were transplanted into GDM rats via tail vein.Blood glucose was measured by blood glucose meter,serum insulin levels were measured before and after transplantation by Elisa method,and the morphology of pancreatic tissue was observed by staining.The blood glucose levels of pregnant rats induced by the cell intervention group were all decreased.It was evident that HE staining induced significant repair of pancreatic tissue in the intervention group. |
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