| 小鼠Islet-1基因慢病毒表达载体的构建及其诱导C_3H_(10)T1/2细胞向心肌样细胞特异性分化 |
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| 引用本文： | 智深深,田杰,刘官信,鲁荣,林建萍,刘建平,朱静. 小鼠Islet-1基因慢病毒表达载体的构建及其诱导C_3H_(10)T1/2细胞向心肌样细胞特异性分化[J]. 基础医学与临床, 2011, 31(7) |
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| 作者姓名： | 智深深 田杰 刘官信 鲁荣 林建萍 刘建平 朱静 |
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| 作者单位： | 1. 重庆医科大学,儿童医院,儿童发育疾病研究省部共建教育部重点实验室,重庆,400014
2. 重庆医科大学,儿童医院,心血管内科,重庆,400014 |
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| 基金项目： | 国家自然科学基金(30973219); 自然科学基金重点项目(CSTC,2009BA5084) |
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| 摘 要： | 目的研究Islet-1对干细胞分化的影响。方法用PCR钓取目的基因,将目的基因与pLenO-WPI载体连接,选取阳性质粒,与辅助质粒共同感染293T细胞生产出慢病毒载体。感染C3H10T1/2细胞,实时荧光定量PCR及Western blot检测Islet-1和心肌、肝脏、骨骼及神经各系统相关标志物的表达,免疫荧光检测心肌肌钙蛋白T(cTnT)表达部位。结果 PCR及测序显示目的片段正确插入,实验组有Islet-1表达;心肌早期发育相关基因GATA-4、MEF2C、NKx2.5在检测到荧光蛋白1周后升高,2周到达高峰,3周后可检测到心肌特异性蛋白cTnT(0.582±0.0576),其时序性表达呈随时间增强趋势;cTnT表达于胞质;肝脏系统特异性标志AFP及ALB、骨骼系统特异性标志BGP及BALP、神经系统特异性标志Nestin及GFAP均未表达。结论 Islet-1具有特异性促进干细胞向心肌样细胞分化的作用。
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| 关 键 词： | 慢病毒表达载体 Islet-1 干细胞特异性分化 心肌样细胞 |
Construction of lentiviral expression vector expressing mouse Islet-1 gene and investigation of Islet-1 mediated induction of C_3H_(10)T1/2 cell differentiation into cardiomyocyte-like cells |
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| ZHI Shen-shen,TIAN Jie,LIU Guan-xin,LU Rong,LIN Jian-ping,LIU Jian-ping,ZHU Jing. Construction of lentiviral expression vector expressing mouse Islet-1 gene and investigation of Islet-1 mediated induction of C_3H_(10)T1/2 cell differentiation into cardiomyocyte-like cells[J]. Basic Medical Sciences and Clinics, 2011, 31(7) |
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| Authors: | ZHI Shen-shen TIAN Jie LIU Guan-xin LU Rong LIN Jian-ping LIU Jian-ping ZHU Jing |
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| Affiliation: | ZHI Shen-shen1,TIAN Jie2,LIU Guan-xin1,LU Rong1,LIN Jian-ping1,LIU Jian-ping1,ZHU Jing1(1.Key Laboratory of Developmental Diseases in Childhood Ministry of Education,2.Dept.of Cardiology,Children's Hospital,Chongqing Medical University,Chongqing 400014,China) |
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| Abstract: | Objective To construct lentiviral expression vectors targeting mouse gene Islet-1 and to study the effect of Islet-1 on stem cell differentiation.Methods Islet-1 gene was obtained by PCR and inserted into pLenO-WPI Vector.The positive plasmid was selected and infected 293T cells with helper plasmid to produce recombinact lentivirus.Islet-1 expression and its related gene,gene and protein markers of heart,liver,bone,and nerves systems were measured by Real-time quantitative PCR and Western blot after C3H10T1... |
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| Keywords: | lentiviral expression vector Islet-1 MSCS differentiation cardiomyocyte-like cells |
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