动物造模,骨髓间充质干细胞 z-bio 2022-06-08 226

　　Objective: To study the ability of adipose stem cells (ASCs) to differentiate into endothelial cells and compare with bone marrow mesenchymal stem cells (BMSCs).

　　METHODS: ASCs and BMSCs were isolated and cultured from SD rats, and well-growing cells of a certain passage were selected, and cell surface markers were identified by flow cytometry; cell growth curves were drawn by CCK-8 method; The expression of endothelial cell specific markers CD31, KDR and vWF mRNA was detected by qPCR; the expression of surface antigen CD31 was observed by immunofluorescence staining; the uptake of induced group cells was detected by Dil-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) Function; Validation of tube-forming ability of induced group cells on Matrigel.

　　RESULTS: Rat ASCs and BMSCs were successfully isolated and cultured. In terms of morphology, ASCs and BMSCs were similar in shape to long spindle, spindle, and fibroblast-like. The results of flow cytometry showed that both BMSCs and ASCs highly expressed mesenchymal stem cells. Surface markers CD29 (100% and 96.9%), CD90 (96.5% and 99.2%), low expression of hematopoietic cell surface marker CD45 (3.9% and 0.8%), confirmed that the extracted mesenchymal stem cells; CCK- 8 The results of proliferation experiments showed that the proliferation rate of ASCs was faster than that of BMSCs (P<0.05). After standard endothelial induction, qPCR showed that the BMSCs and ASCs in the induced group expressed higher levels of CD31, KDR, and vWF mRNA than in the uninduced group (P<0.05). Immunofluorescence staining showed that CD31 antigen was expressed on the cell membrane surface of the induced group. The cells in the group took up Dil-Ac-LDL (the uninduced group did not take up); the cells in the induced group all had the ability to form tubes on Matrigel, which confirmed that the induced cells were endothelial cells.

　　Conclusion: Both BMSCs and ASCs can be induced to differentiate into endothelial cells, and ASCs have a shorter time to extensively differentiate into endothelial cells and have stronger proliferation ability, suggesting that ASCs have more application prospects than BMSCs.