Neural differentiation of allogenic mixed-cultured rat bone mesenchymal stem cells

Many studies showed that bone mesenchymal stem cells (BMSCs) can transdifferentiate to neural cell in vitro. The purpose of this study was to investigate the mixed-culture of allogenic rat BMSCs in vitro, and to perform neural differentiation potential characterization. Rat BMSCs were isolated and cultured by plastic adherence and density gradient centrifugation respectively, and the 3rd passage cells were harvested and mixed-cultured with same concentration. The second passage cells of the mixed-cultured cells were obtained to perform Wright-Gemsa staining for morphological observation, to identify the surface marker of CD29, CD45, CD90 by flow cytometry and induced to differentiate into neural cell, then performed immunocytochemistry of Nestin, NSE and GFAP. Results showed that after Wright-Giemsa staining, the mixed-cultured cells displayed typical spindle-shape, presented layered and whirlpool-like growth; and the mixed-cultured cells were positive for CD29 and CD90, but negative for CD45 by flow cytometry. After induction, the mixed-cultured cells appeared morphological changes of neuron and glial cell, and were positive expression of Nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP). The results demonstrated that the mixed-cultured allogenic rat BMSCs can be induced into neural cell such as neuron and glial cell in vitro.