Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes.
Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes.
复制标题
使用电穿孔将生物活性外源基因引入原代大鼠肝细胞。
DOI：
10.1128/mcb.6.2.716-718.1986
复制
发表时间：
1986
影响因子：
5.3
通讯作者：
Shafritz,DA
中科院分区：
文献类型：
--
作者：
Tur-Kaspa,R;Teicher,L;Levine,BJ;Skoultchi,AI;Shafritz,DA
本文描述了一种在分离肝细胞中引入和表达克隆基因的方法。用pSV2CAT质粒电穿孔法转染经胶原酶灌注分离的大鼠原代肝细胞。48小时后，转染肝细胞的可溶性提取物显示氯霉素乙酰转移酶活性与磷酸钙或deae -葡聚糖转染大鼠肝癌细胞系H4AzC2的活性相当。后两种方法由于具有细胞毒性，不能成功地用于原代肝细胞。这表明电穿孔是获得外源基因在原代上皮细胞（如大鼠肝细胞）中瞬时表达的一种有效方法，这种方法在细胞培养中难以维持。
A method is described for introducing and expressing cloned genes in isolated hepatocytes. Primary rat hepatocytes isolated by collagenase perfusion were transfected in suspension with plasmid pSV2CAT by electroporation. Forty-eight hours later, soluble extracts from transfected hepatocytes showed chloramphenicol acetyltransferase activity comparable to that obtained in rat hepatoma cell line H4AzC2 by calcium phosphate or DEAE-dextran transfection. The latter two methods could not be used successfully for primary hepatocytes because of cytotoxicity of these reagents. This indicates that electroporation is a useful method to obtain transient expression of foreign genes in primary epithelial cells, such as rat hepatocytes, which are difficult to maintain in cell culture.