An improved method for isolation of human umbilical cord mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.10.021

Abstract

Abstract:

BACKGROUND: Human umbilical cord mesenchymal stem cells with capabilities for self-renewal and multi-differentiation have attracted widespread attention.
OBJECTIVE: To develop an efficient method for isolation and culture of human umbilical cord mesenchymal stem cells, and to analyze the cell biological features.
METHODS: Mesenchymal stem cells were isolated and cultured from human umbilical cord by improved tissue cultivation. Immunophenotype and cell cycle were analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated by in vitro osteogenic and adipogenic induction as well.
RESULTS AND CONCLUSION: Some fusiform cells crawled out from human umbilical cord tissues after cultivation for 5 days and formed colonies about 10 days later. When the removed tissues were further cultured, more cells appeared again within 2 days and formed colonies after 5 days. The isolated cells exhibited similar morphology of fibroblast-like shape after passage. Furthermore, the cells expressed CD90, CD105, but were negative for the markers of CD34, CD45, HLA-DR. Population doubling time of the cells calculated from the result of MTT was about 50 hours and cell cycle analysis showed that 41.24% cells were in the G₂/S phrase. Therefore, the isolated cells had a high prolification ability. In addition, the isolated cells could be induced into osteoblasts
and adipocytes in vitro. In a word, the results of this study demonstrated that the cells from the second tissues culture possessed the biological characteristics of mesenchymal stem cells and more primary umbilical cord mesenchymal stem cells were acquired through the improved method.

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: stem cells, umbilical cord, mesenchymal stem cells, cell culture techniques

CLC Number:

R394.2

Li Yan-qi, Wang Hong-yi, Yao Yao, Liu Jing-jing, Xu Xiao, Zhang Yu, Liu Yang, Wu Chu-tse,Jin Ji-de. An improved method for isolation of human umbilical cord mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(10): 1609-1614.

Figures/Tables 5

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