仔猪神经干细胞体外培养及鉴定

由仔猪脑部海马体齿状回分离得到神经干细胞,利用无血清培养基添加特定生长因子方式培养;采用RT-PCR鉴定神经干细胞和诱导分化后细胞生物特性;利用免疫荧光化学技术检测克隆的细胞抗原和分化后特异性成熟神经细胞抗原表达。由海马体齿状回分离的细胞群具有连续克隆能力;根据免疫荧光检测,脑神经干细胞表达巢蛋白、配对盒因子6和解整合素样金属蛋白酶10;体外培养下NSCs可被诱导分化为神经源性细胞,且表达微管相关蛋白2、胶质纤维酸性蛋白和髓鞘碱性蛋白。分离和培育NSCs细胞结果显示,其具有更新能力和分化为神经元和胶质细胞的潜能,可为干细胞移植提供基础数据。

Isolation and characterization of neural stem cells derived from piglets in vitro

Neural stem cells were isolated from the hippocampus dentate gyrus of piglet and cultured with serum-free medium and specific growth factors. Biological characteristics of neural stem cells and induced differentiated cells were identified by RT-PCR. Immunofluorescence cytochemistry technology was used to detect the antigens expression of cloned cells and differentiated mature nerve cells. The cells isolated from the hippocampal dentate gyrus had the ability of continuous cloning.According to immunofluorescence staining detection, Nestin, paired box protein 6 and a disintegrin and metalloprotease 10 were expressed in NSCs. In vitro, NSCs could be induced to differentiate into neurogenic cells and express glial fibrillary acidic protein, microtubule-associated protein 2 and myelin basic protein. NSCs not only displayed a self-renewal capacity, but also had the potential to differentiate into neurons and glial cells, which provided basic data for stem cell transplantation.