Isolation，Culture and Characterization of Derived Cells from Neonatal Porcine Bone Marrow Mesenchymal Stem Cells

doi:10.13560/j.cnki.biotech.bull.1985.2015.06.027

Abstract

Abstract: Recently it has become possible to use bone marrow mesenchymal stem cells(BMSCs)in the clinical treatment of certain diseases. However, BMSCs, the seed cells used in transplantation, are limited in vitro proliferation capability and can only be obtained from very few sources. In order to acquire a substitute for BMSCs, the method of differential velocity adherent screening was used in this study to isolate a derived strain of porcine BMSCs, named bone marrow mesenchymal stem-derived cells(BMSDCs). The biological characteristics of BMSDCs and BMSCs cells were compared by continuous morphological imaging by inverted microscope, and the growth curves of both cells were monitored by the MTT method. Furthermore, their characteristics of differentiation in vitro were examined, and flow cytometry was used to measure cell surface markers. Our results suggest that the doubling times of BMSCs and BMSDCs are 31. 3 h and 30. 3 h, respectively, and the average passaging time is 3-5 d and 2-3 d, respectively. Cultured BMSCs and BMSDCs are both positive for CD34 and CD90 and negative for CD44 and CD45. Both BMSCs and BMSDCs can differentiate into adipocytes and myoblasts by induced differentiation in vitro. Concerning their passaging abilities, the former can have 15 to 20 passages in vitro, whereas latter can have a longer period of time(more than 200 passages)while normal karyotypes are still maintained. We conclude that, under certain experimental circumstances, cultured porcine BMSDCs in vitro can survive and proliferate while maintaining multi-directional differentiation potential of BMSCs, and potentially it can be an ideal type of seed cell for tissue engineering.

Key words: bone marrow mesenchymal stem cell, derived cell, porcine, biological characteristics, tissue engineering

Ruan Zheng, Wang Lianfang, Hu Xiuzhong, Wu Jianying, Zhang Sihua, Dai Changyun, Hua Juan, Xia Yu, Hu Xiaoming, Li Jie, Huang Haijun. Isolation，Culture and Characterization of Derived Cells from Neonatal Porcine Bone Marrow Mesenchymal Stem Cells[J]. Biotechnology Bulletin, 2015, 31(6): 170-176.

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