动物造模,脂肪干细胞 z-bio 2021-12-06 951

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

　　Methods: Separate and culture ASCs and BMSCs from SD rats, select well-growing cells of a certain number of generations, and identify cell surface markers by flow cytometry; draw cell growth curves by CCK-8 method; perform standard endothelial induction for 3 weeks, and use qPCR detection of endothelial cell specific markers CD31, KDR, vWF mRNA expression; immunofluorescence staining to observe the expression of surface antigen CD31; Dil-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) was used to detect the uptake of induced group cells Function; verify the tube-forming ability of the induced group cells on Matrigel.

　　Results: The rat ASCs and BMSCs were successfully isolated and cultured. In terms of morphology, ASCs and BMSCs were similar in shape, both exhibited a long fusiform, spindle-like, and fibroblast-like morphology; 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 proliferation experiment results showed that the proliferation rate of ASCs was faster than that of BMSCs (P<0.05). After standard endothelial induction, qPCR showed that the expression of CD31, KDR, and vWF mRNA in BMSCs and ASCs in the induced group was higher than that in the uninduced group (P<0.05); immunofluorescence staining showed that CD31 antigen was expressed on the cell membrane surface of the induced group; induced under a fluorescence microscope Cells in the group uptake Dil-Ac-LDL (no uptake in the uninduced group); cells in the induced group have the ability to tube on Matrigel, confirming that the induced cells are endothelial cells.

　　Conclusion: It shows that both BMSCs and ASCs can induce differentiation into endothelial cells, and the extensive differentiation of ASCs into endothelial cells is shorter and the proliferation ability is stronger, suggesting that ASCs has more application prospects than BMSCs.