【摘要】 目的　建立自体肾系膜细胞作为靶向载体转导外源基因的方法 ,使组织金属蛋白酶抑制剂 1基因在肾系膜细胞稳定表达。方法　选择性DMEM培养液培养的肾活检组织系膜细胞 ,构建重组真核表达载体pcDNA3-TIMPl进行体外转染 ,G4 18筛选阳性克隆 ,RT -PCR确认。结果　建立了人肾穿刺组织系膜细胞培养方法 ,经组织学和免疫组化鉴定系膜细胞 ,成功构建表达组织金属蛋白酶抑制剂 1基因的重组真核表达载体pcDNA3-TIMP1,筛选得到一个稳定表达的阳性克隆。结论　肾活检组织系膜细胞培养条件的建立为肾病的基因治疗提供了一种有效载体 ,稳定表达组织金属蛋白酶抑制剂 1基因的肾系膜细胞株为研究TIMPl在肾脏病中的作用提供良好的模型。更多还原

【Abstract】 Objective:To study the suitable condition to culture autologous mesangial cellsfrom renal biopsy specimens for gene delivery. To constitute the cell strain stably express TIPM1 gene.Methods:Mesangial cells of 5 specimens renal biopsy were cultured with selectiveDMEM culture liquid. TMIP 1 eDNA was amplified for c onstructing the plasmid pcDNA3-TMIP 1,which was transfected into MsC. G418 scann ing and RT-PCR were used to identify the positivecell clone.Results:Method of mesangial cells cultured in vitro was established. These cells wer e identified for human MC with microscopical observations and immunohistochemica l method. Onepositive clone expressing TMIP1 gene stably was obtained .C onclusions:The success culture ofautologous mesangial cells can provide gene vector and the TIMP1 positive clone is valuable for providing experimental bases of the nephropathy genetic therapy.更多还原