Empirical studying of bone marrow mesenchymal stem cells transdifferentiated into islet-like cells in vitro
AIM: Rat Bone marrow mesenchymal stem cells (BMSCs) were induced into islet-like cells in vitro by different dosage glucose culture medium, then observed the differences of induced cells. METHODS: To verify the morphologic changes of the cells by Invert microscope. The insulin-like cells were identified by dithizone (DTZ) and immunocytochemistry. TO estimated the reaction of induced cells by radio-immunity analysis (RIA) after glucose stimulation. The expression levels of gene about the induced group cells and control group cells were measured by realtime quantitative PCR. RESULTS: Comparing the non-induced with the transdifferentiated ceils uhramicrostructure by electron microscope, some changes were proposed. Insulin in the induced ceils was examined by DTZ stain. To observe the morphology using microscope, the induced group ceils were stained and the control group ,lot. Immunocytochemical staining showed that low-glucose induced group A value of (4.75 ± 1.87 ), mid-glucose induced group A value of( 10.96 +2.06), high-glucose induced group A value of( 12.53 ± 2.04), as the induced group glucose concentration increased, integral optical density values rising. Real-time PCR quantitative analysis showed that pancreatic 13-cell-related genes GK, PDX-1, ngn3, GLP-1 Mrna expression level after induction has changed significantly, the difference was significant (P <0.01 ). Under high glucose stimulation , insulin secretion in high glucose-induced cells was(6.64 ±0.46) IU/L by RIA, higher than the other groups, the difference was significant ( P < 0. 01 ).CONCLUSION: The rat BMSCs may be induced to differentiate into islet-like cells in different dosage glucose culture medium. After a short period of observation, the effect of high-glucose induced group is superior to low-glucose induced group and midglucose induced group.
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