TGF-β1-induced Differentiation of Wapiti Antler Mesenchymal Stem Cells to Cartilage and Expression Profile of Gene c-myc

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.018

Abstract

Abstract: Antler mesenchymal stem cells(MSCs)play a pivotal role on the antler regeneration and ossification. To investigate the chondrogenic differentiation of antler MSCs and the regulation role of the proto-oncogene c-myc in this process, the second passage cells(MSCs, P2)of 60 d antler from adult Tarim wapiti were induced to chondrogenesis by the stimulation of transforming growth factor TGF-β1(10 ng/mL)in vitro. The inducing effects were identified by Alcian blue staining and immunohistochemics, and the expressions of gene c-myc during this process were detected by qPCR. The results demonstrated that, on the day 9 after induction, some MSCs begun to change from spindle-shaped to rounded or polygon, and the chrysanthemums-shaped pattern of the original MSCs gradually changed to paving stone like；the cartilage capsules were observed on the day 14, and the cartilage extracellular matrix was obvious and cell apoptosis appeared on the day 21. While the cells in the control group were observed to have apoptosis on the day 28, and the cavitations were discovered in the cells. On the day 35, massive cell apoptosis of both groups were observed, and the cell refraction became weak and the gaps between cells became larger. The identification by Alcian blue staining revealed that heavy positive staining emerged in the cellular matrix from the day 14 after stimulating. The detection by immunohistochemistry demonstrated that positive brown Col II reactant was in the cellular matrix from the day 21 after stimulating, and the color became darker along with the culture time, mainly distributed in the cells and their surround matrix. In the cartilage differentiation process from the day 7 to 28, the expressions of gene c-myc in the cells of the induced group were significantly lower than that of control group(P<0.05), while there was no significant difference after the day 35(P>0.05). In conclusion, Tarim wapiti antler MSCs differentiated into cartilage under the stimulation of TGF-β1. The down-regulated expression of proto-oncogene c-myc induced the apoptosis of antler MSCs and then differentiation to chondrogenic cells.

Key words: wapiti antler, mesenchymal stem cells, differentiation, transforming growth factor β1, c-myc gene

HAN Chun-mei, WANG Shan-shan, GAO Qing-hua, ZHENG Yong-fu, MA Meng-ting, ZHANG Qin. TGF-β1-induced Differentiation of Wapiti Antler Mesenchymal Stem Cells to Cartilage and Expression Profile of Gene c-myc[J]. Biotechnology Bulletin, 2016, 32(3): 109-114.

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