In vitro osteogenic differentiation and identification of rabbit bone marrow mesenchymal stem cells isolated and cultured with whole bone marrow adherent culture method

doi:10.3969/j.issn.2095-4344.2013.06.020

Abstract

Abstract:

BACKGROUND: The flow cytometry and immunomagnetic separation method have greater impact on cell viability, and although the density gradient centrifugation method can be used to obtain the high purity monocytes, however, the multiple centrifugation may cause a huge loss of cells and has some impact on the cell activity, so the application is debatable.
OBJECTIVE: To isolate and culture rabbit bone marrow mesenchymal stem cells with whole bone marrow adherent culture method for osteogenic differentiation and identification.
METHODS: The rabbit bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adherent culture method, then the morphological characteristics of rabbit bone marrow mesenchymal stem cells were observed under inverted microscope. Under the induction of osteogenic inducer, alkaline phosphatase staining, typeⅠcollage immunocytochemical staining, alizarin red staining and Von-Kossa staining were performed to observe the morphology of rabbit bone marrow mesenchymal stem cells after osteogenic induction by electron microscope.
RESULTS AND CONCLUSION: After in vitro induced differentiation, rabbit bone marrow mesenchymal stem cells exhibited the morphological and biological characteristics similar to typical osteoblasts, alkaline phosphatase staining, type Ⅰ collagen immunocytochemical staining, and Von-Kossa staining and alizarin red mineralization nodules staining were positive. All results indicate that rabbit bone marrow mesenchymal stem cells in vitro isolated and cultured with whole bone marrow adherent culture method can be induced to differentiate into osteoblasts after osteogenic induction.

Key words: stem cells, stem cell culture and differentiation, whole bone marrow adherent culture method, bone marrow mesenchymal stem cells, cell culture, osteogenic differentiation, tissue-engineering bone, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Xiao Shi-hui, Wei Qing-jun, Zhao Jin-min, Bo Zhan-dong, Wei Ji-hua, Li Wei-an. In vitro osteogenic differentiation and identification of rabbit bone marrow mesenchymal stem cells isolated and cultured with whole bone marrow adherent culture method[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(6): 1069-1074.

Figures/Tables 4

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