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Study of Rabbit Bone Marrow Mesenchymal Stem Cells Differentiating into Islet-Like Cells by DMSO Combined with High Glucose In Vitro
Yu Wenhao, Wang Yadan, Hu Xiao, Zhang Ning, Xue Yongkang, Zhu Hanyu, Zhao Yuxi, Wang Xinzhuang*
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China
Abstract: The aim of this study was to explore the feasibility and regulation mechanism of rabbit bone marrow mesenchymal stem cells (BMSCs) differentiating into islet-like cells. Rabbit BMSCs were inducted into islet-like cells by dimethyl sulphoxide (DMSO) combined with high glucose and without serum. The morphological changes of cells were observed under the inverted microscope. The differentiated cells were detected by dithizone (DTZ) staining and immunofluorescence staining. The changes of the expression of islet cell related genes [Foxa2 (forkhead box A2)， Nestin (neuroepithelial stem cell protein)， Pax6 (paired box gene 6)， Pdx-1 (pancreatic duodenal homeobox-1)， Insulin] were detected by RT-qPCR， and the expression of islet cell related genes in cells before induction (0 d)， rabbit bonemarrow cells， P3 BMSCs and rabbit pancreatic tissues were used as references. The results showed that Foxa2， Nestin and Pax6 genes couldnot be used as the marker genes for rabbit BMSCs differentiating into islet-like cells successfully. DMSO could activate the expression of Pdx-1 and promote rabbit BMSCs to differentiate into islet precursor cells which can secrete insulin. High glucose could promote rabbit BMSCs to differentiate into islet-like cells and could significantly promote the expression of Pdx-1 and Foxa2 genes.
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