Culture of bone marrow mesenchymal stem cells in vitro and effect of hypoxia on bone marrow mesenchymal stem cells

Abstract

Objective To isolate, culture and identify rat bone marrow mesenchymal stem cell (BMMSC) in vitro, and observe the effect of hypoxia on proliferation of BMMSC. Methods Adult male SD rats were sacrificed by decapitation, and soaked in 75% alcohol for five minutes.Cells were then cultured by the whole bone marrow adherent method, and the third generation cells in good condition were selected for identification.The rat BMMSC was identified by flow cytometry.CD90, specific surface marker of BMMSC, and CD45, nonspecific surface marker of BMMSC, were chosen to be detected to identify BMMSC.The proliferation of the third generation cells and cells treated by hypoxia was determined by MTT. Results BMMSC could be isolated and cultured from SD rats.Results of flow cytometric test showed that the expression rate of CD90 was 96.0%, while the expression rate of CD45 was only 2.5%.MTT results indicated that the proliferation rate of BMMSC under hypoxic training was higher than that under normal incubation condition.In addition, the cell state and refraction were also better than those under the normal condition. Conclusions BMMSC can be isolated and cultured in vitro from SD rats by the whole bone marrow adherence method, and hypoxia can stimulate the proliferation of BMMSC. Key words: Mesenchymal stem cell; Bone marrow; Cell culture; Cell