Setting optimal parameters for in vitro electrotransfection of B16F1, SA1, LPB, SCK, L929 and CHO cells using predefined exponentially decaying electric pulses
Setting optimal parameters for in vitro electrotransfection of B16F1, SA1, LPB, SCK, L929 and CHO cells using predefined exponentially decaying electric pulses
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DOI：
10.1016/j.bioelechem.2003.10.009
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发表时间：
2004-04-01
影响因子：
5
通讯作者：
Novakovic, S
中科院分区：
文献类型：
--
作者：
Cegovnik, U;Novakovic, S
为了实现质粒DNA到细胞中的最大引入，同时为了防止不期望的细胞死亡，电转染条件应针对每种单个细胞类型单独确定。在本研究中，我们确定了体外转染B16 F1，SA 1，LPB，SCK，L929和CHO细胞的最佳电转染参数。这些变化的参数中的一些是电场强度、施加的脉冲的数量和它们的持续时间、电穿孔缓冲液的渗透压、质粒DNA浓度和进行电穿孔的温度。在最佳电转染参数下，B16 F1、SA 1、LPB、SCK、L929和CHO的最大转染率分别为85%、40%、60%、1%、40%和65%。所获得的结果证实，电穿孔是一个有用的程序，在体外转染的大多数哺乳动物细胞。如果优化，该方法可以在对电场作用敏感的细胞类型中产生可再现的高比例的转染细胞。因此，所确定的参数可以为该方法的后续实施服务。(C)2003 Elsevier B.V.保留所有权利。
To achieve the maximal introduction of plasmid DNA into cells and, at the same time, to prevent undesirable cell deaths, electrotransfection conditions should be determined for every single cell type individually. In the present study, we determined the optimal electrotransfection parameters for in vitro transfection of B16F1, SA1, LPB, SCK, L929 and CHO cells. Some of these varying parameters were electric field strength, number of applied pulses and their duration, osmolarity of electroporation buffer, plasmid DNA concentration and temperature at which the electroporation was carried out. The maximal transfection rates at optimal electrotransfection parameters in B16F1, SA1, LPB, SCK, L929 and CHO were 85%, 40%, 60%, 1%, 40% and 65%, respectively. The obtained results confirmed that the electroporation is a useful procedure for an in vitro transfection of the majority of mammalian cells. The method, if optimized, may generate reproducibly high proportion of transfected cells among the cell types that are sensitive to electric field action. Thus, the determined parameters could serve for the subsequent implementations of this method. (C) 2003 Elsevier B.V. All rights reserved.