Isolation, culture and CM-Dil labeling of rat mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2014.01.007

Abstract

Abstract:

BACKGROUND: Currently, there is no uniform, standardized approach to isolate, purify and proliferate bone marrow mesenchymal stem cells. Chlormethylbenzamido-1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (CM-Dil) is a stable, reliable, high marking and simple marker.
OBJECTIVE: To develop the methods for isolation, culture and identification of rat bone marrow mesenchymal stem cells in vitro.
METHODS: Two male Sprague-Dawley rats, weighing 50-100 g were taken to collect the bilateral femur and tibia bone marrow under sterile conditions, and then, primary bone marrow mesenchymal stem cells were isolated and cultured using bone marrow adherent separation and density gradient centrifugation. Cells were amplified and purified through timely and repeated passage, and labeled at the third generation with fluorescent dyes CM-Dil in vitro as a source of donor cells.
RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were cultured successfully in vitro using bone marrow adherent separation and density gradient centrifugation separation methods, but the former was superior to the latter in the number of cultured cells significantly, while the two methods were not different significantly in terms of cell viability and proliferation. Flow cytometry results showed that the positive rates of cultured cells were 17.5% for CD34, 97.9% for CD44, and 91% for CD90. CM-Dil can label bone marrow mesenchymal stem cells successfully, which is a stable, reliable, high marking and simple marker.

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, mesenchymal stem cells, cells, cultured, biological markers

CLC Number:

R394.2

Li Chao-zhong, Xiao Jian-ming, Chen Li-xing, Li Wan-rong, Zhang Chun-hai . Isolation, culture and CM-Dil labeling of rat mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(1): 39-44.

Figures/Tables 3

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