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目的探讨人骨髓间充质干细胞(hBMSCs)体外多向分化的潜能,为体内移植实验提供种子细胞。方法采用全骨髓贴壁培养法,分离、自制无血清培养基培养hBMSCs;取生长良好的P4代hBMSCs,免疫荧光法检测其表面抗原CD34、CD44和CD105的表达;应用含不同诱导剂的培养基进行诱导培养,油红O染色检测成脂诱导分化,茜素红染色鉴定成骨细胞诱导分化,阿利新蓝染色检测成软骨诱导情况。结果 hBMSCs表面抗原CD44、CD105呈阳性,CD34呈阴性;经过诱导培养,油红O染色、茜素红染色以及阿利新蓝染色均呈阳性。结论分离培养的hBMSCs能够分化为成脂细胞、成骨细胞和软骨细胞,说明hBMSCs具有多向分化的潜能,可以作为体内移植实验较为理想的种子细胞。
Objective To explore the potential of multidirectional differentiation of human bone marrow mesenchymal stem cells (hBMSCs) in vitro and to provide seed cells for in vivo transplantation experiments. Methods The whole bone marrow adherent culture was used to culture hBMSCs isolated and self-made serum-free medium. The well-grown P4 generation hBMSCs were cultured and the expression of surface antigens CD34, CD44 and CD105 were detected by immunofluorescence. Induction culture was conducted. Oil red O staining was used to detect adipogenic differentiation. Alizarin red staining was used to identify osteoblasts. Differentiation of osteoblasts was induced by alizarin blue staining. Results The surface antigen CD44 and CD105 of hBMSCs were positive and CD34 was negative. After induced by oil red O staining, alizarin red staining and alitame blue staining, all the hBMSCs were positive. Conclusion The hBMSCs isolated and cultured can differentiate into adipocytes, osteoblasts and chondrocytes, indicating that hBMSCs have the potential of multidirectional differentiation and can be used as ideal seed cells for in vivo transplantation experiments.