Ficoll密度梯度离心法分离培养纯化猪的骨髓间充质干细胞

发布时间：2018-11-01 12:40

【摘要】：背景:研究报道显示猪骨髓间充质干细胞体外分离方法、效率、培养条件各不相同,并且尚无统一的鉴定标准。目的:建立一种高效、经济的猪骨髓间充质干细胞分离培养及鉴定方法。方法:采集健康猪股骨骨髓,Ficoll密度梯度离心法分离单个核细胞,进行原代和传代培养,流式细胞仪检测细胞膜表面抗原表达情况,并观察细胞成骨、成脂及成软骨分化的能力。结果与结论:分离培养的单个核细胞活力旺盛,体外生长良好,细胞呈梭形、纺锤形和小多角形,具有稳定的增殖分裂能力;细胞膜表面抗原CD29、CD44、CD90呈阳性表达,而CD14、CD34、CD45、CD166、HLA-DR呈阴性表达;在特定诱导条件下,该细胞可分化为成骨细胞、脂肪细胞和软骨细胞。说明采用Ficoll密度梯度离心法成功分离培养出纯化的猪骨髓间充质干细胞,其生长状态良好,具有多项分化潜能,可作为组织工程的种子细胞。
[Abstract]:Background: in vitro isolation of porcine bone marrow mesenchymal stem cells (BMSCs) has been reported. Objective: to establish an efficient and economical method for isolation, culture and identification of porcine bone marrow mesenchymal stem cells. Methods: the bone marrow of healthy pig femur was collected and mononuclear cells were isolated by Ficoll density gradient centrifugation. The expression of cell surface antigen was detected by flow cytometry and osteogenesis was observed. The ability to differentiate into fat and cartilage. Results and conclusion: the mononuclear cells isolated and cultured had high activity, good growth in vitro, spindle shape, spindle shape and small polygonal shape, and had stable proliferative and mitotic ability. The cell membrane surface antigen (CD29,CD44,CD90) was expressed positively, but CD14,CD34,CD45,CD166,HLA-DR was negative. Under specific induction conditions, the cell could differentiate into osteoblasts, adipocytes and chondrocytes. The results showed that the purified porcine bone marrow mesenchymal stem cells were successfully isolated and cultured by Ficoll density gradient centrifugation. The MSCs grew well and had multiple differentiation potential, so they could be used as seed cells for tissue engineering.
【作者单位】：解放军第307医院心血管内科;
【分类号】：R329

【共引文献】