Isolation, cultivation and identification of porcine bone marrow mesenchymal stem cells using Ficoll density gradient centrifugation method

doi:10.3969/j.issn.2095-4344.2014.50.006

Abstract

Abstract:

BACKGROUND: Culture condition, isolation method and efficiency of porcine bone marrow mesenchymal stem cells are different, and there is a lack of unified identification standards.
OBJECTIVE: To establish a high-efficiency and economical method of isolating, culturing and identifying porcine bone marrow mesenchymal stem cells.
METHODS: The bone marrow was collected from healthy porcine femur to isolate mononuclear cells by Ficoll density gradient centrifugation method. After primary and subculture, the cell surface antigens were detected by flow cytometery, and the osteogenic, adipogenic and chondrogenic differentiation abilities of the cells were observed.
RESULTS AND CONCLUSION: The isolated mononuclear cells showed exuberant viability, and grew well in vitro. They were spindle- or small polygonal-shaped and had a stable doubling time on culture. Flow cytometric analysis revealed that they were positive for CD29, CD44, CD90, and negative for CD14, CD34, CD45, CD166, HLA-DR. Furthermore, they were able to differentiate into osteogenic, adipogenic and chondrogenic lineages under specific conditions in vitro. Porcine bone marrow mesenchymal stem cells isolated by Ficoll density gradient centrifugation method can be purified, grow well in vitro, and have multi-lineage differentiation potentials, which can act as seed cells for tissue engineering research.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: bone marrow, mesenchymal stem cells, cell culture techniques, cells, cultured, swine

CLC Number:

R394.2

Liu Chuan-bin, Lǚ Shuang-hong, Zhang Xiao-zhong. Isolation, cultivation and identification of porcine bone marrow mesenchymal stem cells using Ficoll density gradient centrifugation method[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(50): 8066-8071.

Figures/Tables 2

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