Isolation, culture and identification of SD rat bone marrow mesenchymal stem cells in tissue engineering

Abstract

Seed cells are an important part of bone tissue engineering. In this study, we investigated methods for the isolation, culture and identification of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. BMSCs from SD rats were isolated and cultured using the whole bone marrow adherent method. Morphological observation and flow cytometry were used for the detection of cell surface markers. The osteogenic and adipogenic differentiation of cultured cells was induced and evaluated. BMSCs could be effectively isolated and purified using the whole bone marrow adherent method. The primary BMSCs cultured in vitro formed spindle-shaped, spiral and vigorous cell colonies. The flow cytometry results showed that CD90 was highly expressed on the cell surface, but CD34 and CD45 were not expressed. After culture in osteogenic and adipogenic induction medium, positive Alizarin Red and Oil Red O staining was observed, indicating the potential for multidirectional differentiation. The whole bone marrow adherent method is simple and practical, which makes the isolation, culture and amplification of BMSCs in vitro easy, and the method has little effect on cell activity and is an effective method for obtaining high-purity BMSCs.