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首页> 外文期刊>Molecular and Cellular Biology >Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes.
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Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes.

机译：使用电穿孔将生物活性外源基因引入原代大鼠肝细胞。

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摘要

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.
机译：描述了在分离的肝细胞中引入和表达克隆基因的方法。通过电穿孔将通过胶原酶灌注分离的原代大鼠肝细胞悬浮于质粒pSV2CAT中。 48小时后，从转染的肝细胞中提取的可溶性提取物显示出氯霉素乙酰转移酶活性，与通过磷酸钙或DEAE-葡聚糖转染在大鼠肝癌细胞系H4AzC2中获得的活性相当。由于这两种试剂的细胞毒性，后两种方法不能成功用于原代肝细胞。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）中瞬时表达的有用方法，而原代上皮细胞难以在细胞培养物中维持。

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