OALib Journal期刊
ISSN: 2333-9721
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人神经营养因子-3基因的亚克隆及其基因工程细胞模型的建立
, PP. 805-808
Keywords: 神经营养因子-3/分泌,骨髓间充质干细胞/遗传学,转染
Abstract:
目的亚克隆人神经营养因子-3(humanneurotrophin-3,nt3)并转染至体外培养的人骨髓间充质干细胞(bm-mscs),建立能够表达和分泌nt3的基因工程细胞模型。方法采用低糖dmem培养液+10%胎牛血清体外培养bm-mscs,流式细胞仪分析其表面标记;构建真核表达载体pcdna3.1(+)/nt3并通过脂质体介导转染bm-mscs;收集部分细胞提取其中的总rna,经逆转录pcr检测nt3基因表达;利用培养上清液体外孵育毛细胞以鉴定其生物学活性。结果bm-mscs表达cd13、cd29、cd59,不表达cd11、cd14、cd31、cd34、cd45、cd80、cd86、cd117、hla-dr;pcdna3.1(+)/nt3转染bm-mscs后,能够表达和分泌nt3,并具有生物学活性,可促进离体的豚鼠耳蜗毛细胞存活。结论在体外建立表达nt3基因并分泌nt3的基因工程细胞是可行的。
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