胎盘源性间充质干细胞多药耐药基因的转染与鉴定
韩丽英;李亚萍;叶明珠;王博蔚;王强;赵淑华;李荷莲
【期刊名称】《中华医学杂志》 【年(卷),期】2012(092)041
【摘要】目的 探讨重组逆转录病毒载体介导mdr1基因转染胎盘源性间充质干细胞(P-MSC)以应用于基因治疗的可行性、安全性.方法 Percoll密度梯度离心法自人胎盘组织中分离培养间充质干细胞(MSC),利用重复感染方法将含有mdr1基因的逆转录病毒载体导入P-MSC,经逆转录多聚酶链式反应(RT-PCR)检测mdr1 mRNA表达,荧光细胞分选法(FAGS)检测目的基因表达,同时对mdr1-MSC的干细胞特性进行鉴定及分析.结果 转染细胞中显示mdrl mRNA表达；转染P-MSC表达P-gp的阳性细胞百分率为(27.6±5.1)％,明显高于非转染组的(0.4±0.1)％,两组比较差异有统计学意义(t=14.291,P＜0.01)；转染MSC在G0/G1期所占比例为95.40％,符合干细胞增殖特点.超微结构揭示其具有典型的低分化干细胞超微结构特点,且在特异诱导条件下具有向成脂、成骨定向分化潜能.结论 由重组逆转录病毒介导mdr1基因体外转染胎盘源性MSC可获得高效的P-gp表达,且转染细胞可依然保持其干细胞特性.%Objective To explore the feasibility and safety of mdr1 gene transferred into placenta derived mesenchymal stem cells (P-MSCs) by reconstructed retroviral vector.Methods Human P-MSCs were isolated and expanded by Percoll density gradient and then transduced repeatedly by reconstructed retroviral vector containing mdr1 gene.The transfection and expression of mdr1 gene was detected by reverse transcription-polymerase chain
reaction (RT-PCR) and fluorescence-activated cell sorting
(FAGS).Meanwhile,the biological features of mdr1-MSCs were identified and analyzed.Results The expression of mdr1 mRNA was found in transfected cells.The expression of P-glycoprotein (P-gp) encoded by mdr1 gene was(27.6 ±5.1)％ in the transfected P-MSCs cells versus (0.4 ± 0.1)％ in the non-transfected P-MSCs cells (t =14.291,P ＜ 0.01).The percent of P-MSCs at quiescent phase (G0/G1 phase) was around 95.40％ and it was in accord with the characterization of stem cells.The mdr1-MSCs exhibited typical ultrastructures of low-differentiated stem cells.Moreover,they still retained the potency of adipogenic and osteogenic differentiation in the presence of appropriate conditioned media.Conclusion A stable expression of P-gp may be obtained by reconstructed retroviral-mediated transfection in vitro.And transfected MSCs retain the characteristics of stem cells. 【总页数】4页(2930-2933) 【关键词】干细胞;胎盘;转染;基因
【作者】韩丽英;李亚萍;叶明珠;王博蔚;王强;赵淑华;李荷莲
【作者单位】130041长春,吉林大学第二医院妇产科;130041长春,吉林大学第二医院眼科;厦门大学附属中山医院妇产科;130041长春,吉林大学第二医院妇产科;130041长春,吉林大学第二医院妇产科;130041长春,吉林大学第二医院妇产科;130041长春,吉林大学第二医院妇产科 【正文语种】中文
【中图分类】 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_national-medical-journal-china_thesis/0201255079882.html 【相关文献】
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胎盘源性间充质干细胞多药耐药基因的转染与鉴定
