Vascular endothelial growth factor combined with basic fibroblast growth factor improves replicative senescence of bone marrow mesenchymal stem cells

doi:10.12307/2024.199

Abstract

Abstract: BACKGROUND: Mesenchymal stem cells are susceptible to senescence during in vitro expansion, which greatly hinders their application in vivo and in vitro. How to improve the replicative senescence of mesenchymal stem cells is an urgent problem to be solved in tissue engineering.
OBJECTIVE: To determine whether vascular endothelial growth factor combined with basic fibroblast growth factor can improve the aging of bone marrow mesenchymal stem cells caused by replicative passage.
METHODS: Rat bone marrow mesenchymal stem cells were extracted by whole bone marrow adhesion method. Passage 2 cells were selected as normal control group. Passage 7 and later algebraic cells were selected as aging model group. Vascular endothelial growth factor (50 μg/L), basic fibroblast growth factor (10 μg/L), and their combination were administered. Cell proliferation was detected by CCK-8 assay. Cell senescence was observed by β-galactosidase activity staining. Cytoskeleton size and colony formation ability were observed by phalloidine staining and Giemsa staining, respectively, and the levels of senescence-related genes P16, P21, and P53 were detected by qRT-PCR. Gene expression levels of P16, P21, and P53 were tested by qRT-PCR.
RESULTS AND CONCLUSION: (1) Vascular endothelial growth factor combined with basic fibroblast growth factor could promote the proliferation of aged bone marrow mesenchymal stem cells, which began to enter the plateau stage on day 9, and the absorbance value of the combined intervention group was significantly higher than that of the model group on day 9 (P < 0.05). (2) The phenotypic markers of the cells in the combined intervention group did not change, and the cell morphology changed from broad to slender. (3) Compared with the model group, the positive rate of β-galactosidase was significantly decreased (P < 0.01); the number of nuclei increased (P < 0.001); the total area of cytoskeleton increased (P < 0.01); colony formation ability was enhanced (P < 0.05); expression level of P16 was decreased (P < 0.01) in the combined intervention group. These results indicate that vascular endothelial growth factor combined with basic fibroblast growth factor can improve the senescence of bone marrow mesenchymal stem cells caused by replicative passage without changing the cell phenotype.

Key words: vascular endothelial growth factor, basic fibroblast growth factor, bone marrow mesenchymal stem cell, replicative senescence

CLC Number:

Shi Weili, Liu Shanshan, Chang Hongbo, Gao Haixia, Wang Xinzhou, Qin Nan, Wu Hong. Vascular endothelial growth factor combined with basic fibroblast growth factor improves replicative senescence of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(31): 4958-4963.

Figures/Tables 5

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