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目的探讨不同电穿孔条件对原代大鼠肝细胞生物学性状的影响，优化外源基因转染原代肝细胞的条件。方法以低电压 （２２０～４００ Ｖ）、高电容（５００～ ９５０μＦ）单脉冲电击原代大鼠肝细胞后接种培养，分别于 ３６ ｈ～９ｄ不同时点采用台盼蓝拒染法与 ＭＴＴ法观测肝细胞活力，并检测肝细胞培养上清液中白蛋白（ＡＬＢ）、ＡＬＴ与乳酸脱氢酶 （ＬＤＨ）含量。以未电穿孔肝细胞为对照组。结果培养 ３６ ｈ～９ ｄ各组肝细胞存活率均高于９０％，贴壁率高于９５％。培养３６ ｈ肝细胞上清液中ＡＬＢ、ＡＬＴ与ＬＤＨ值，电穿孔Ｂ组（２２０ Ｖ，９５０μＦ）、Ｃ组（４００ Ｖ， ９５０μＦ）明显高于对照组；但 ＭＴＴ法检测各电穿孔组肝细胞活力与对照组比较差异无显著性。结论该电穿孔条件不影响肝细胞的存活能力，还能促进肝细胞ＡＬＢ等蛋白的合成与释放，可作为外源基因转染原代肝细胞的优选方法之一。
Objective To investigate the effects of different electroporation conditions on the biological characteristics of primary rat hepatocytes and to optimize the conditions for transfecting primary hepatocytes with exogenous genes. Methods Primary rat hepatocytes were shocked with low voltage (220-400 V) and high capacitance (500-950 μF) by single pulse. The primary cultured rat hepatocytes were stained with trypan blue exclusion method and MTT method at different time points from 36 h to 9 d, The viability of hepatocytes was observed. The levels of albumin (ALB), ALT and lactate dehydrogenase (LDH) in hepatocyte culture supernatants were detected. Electroporated hepatocytes were used as control group. Results The survival rates of hepatocytes in all groups from 36 h to 9 d were all higher than 90%, and the attachment rate was higher than 95%. The ALB, ALT and LDH values in 36 h hepatocyte supernatants were significantly higher in electroporation group B (220 V, 950 μF) and C group (400 V, 950 μF) than those in control group. However, Cell viability was not significantly different from the control group. Conclusion The electroporation condition does not affect the viability of hepatocytes, but also promotes the synthesis and release of ALB and other proteins in hepatocytes, which can be used as one of the preferred methods for transfection of primary hepatocytes with exogenous genes.