The Study Of Biomarkers Of Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells And Differentiated Into Insulin-producing Cells By Using High-resolution Magic Angle Spinning Magnetic Resonance Spectroscopy

Objective:To study the biomarkers of Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells (huMSCs) and insulin-producing cells differentiation of huMSCs by using in vitro 9.4T high resolution magnetic resonance (MR) spectroscopy, to find huMSCs and insulin-producing cells differentiation of huMSCs'characterization metabolites, identification and tracking the process of differentiation.Methods:Isolating culture collection of births preterm health cesarean section umbilical cord and cultured huMSCs and induced huMSCs differentiated into insulin-producing cells in the condition of islet cells grows(H-DMEM contains 0.1mmol/L Beta mercaptoacetic ethanol,10ug/L bFGF,10ug/L EGF induces stem cells to appear changes,then use the H-DMEM contains 10mmol/L niacinamide to induce about 6 days). The morphology of huMSCs after induction were monitored by under inversion phase contrast microscope; Dithizon-stained was used to detect zinc hydronium and expresses proof differentiation model success.1H-data acquisitions were performed by using in vitro 9.4T high resolution MR spectroscopy (Bruker Avance 400MHz). Spectra was primarily processed in the Frequecvy domain with the XWINNMR (Bruker GmBH) and then analyzed with Mestre 4.7.Results:The morphology of huMSCs under medicine induction gradually changed from fibroblast to round and some of then had the tend of forming clusters.;Dithizon stain show that the cytoplasm of huMSCs after induction were stained in Brownish red color;the spectrums of the huMSCs were high quality and good reproducibility. Several major metabolites can be observed in the huMSCs spectra, includingcholine compound, glutamine, acetic acid, lysine, leucine, isoleucine, valine, alanine. The differentiations of metabolites between the huMSCs and the huMSCs after medicine induction are Isoleucine, valine, lysine increased from3.44±0.54,0.65±0.14,0.16±0.074 nmM to 4.66±0.42(p<0.05),1.05±0.21(p<0.05),0.31±0.052 nmM(p<0.05); Glutamate declined, 0.17±0.058 nmM to 0.064±0.089 nmM(p<0.05), and alanine and leucine had no change, myo-Inositol appear in the spectra of medicine induce huMSCs.Conclusion:The spectrums of the huMSCs were high quality and good reproducibility. According to the differentiations of metabolites change, MR spectroscopy is capable of monitoring the cells of medicine induced huMSCs to insulin-producing cells.