通过快速冷冻将基因导入哺乳动物细胞。
Gene transfer into mammalian cells by rapid freezing.
作者信息
Sasaki K, Mizusawa H, Ishidate M, Tanaka N
机构信息
Division of Cell Biology, Hatano Research Institute, Food and Drug Safety Center, Kanagawa, Japan.
出版信息
In Vitro Cell Dev Biol. 1991 Jan;27(1):86-8. doi: 10.1007/BF02630898.
In this paper, we describe a simple technique to introduce DNA into cells through cracks and/or pores in cell membranes caused by intracellular ice crystal formation induced by liquid nitrogen. We mixed mouse BALB 3T3 cells and pSV2-neo DNA and froze the cell suspension under various conditions to determine those optimum for the introduction of DNA into mammalian cells. We found that brief treatment with liquid nitrogen, which showed only moderate cell killing, resulted in the induction of G-418 resistant colonies. These results suggest that this new technique is useful for transfection of genes into mammalian cells.
在本文中，我们描述了一种简单的技术，可通过液氮诱导细胞内形成冰晶而导致细胞膜出现裂缝和/或孔隙，从而将DNA导入细胞。我们将小鼠BALB 3T3细胞与pSV2-neo DNA混合，并在各种条件下冷冻细胞悬液，以确定将DNA导入哺乳动物细胞的最佳条件。我们发现，用液氮进行短暂处理，虽然细胞杀伤程度适中，但能诱导出对G-418具有抗性的菌落。这些结果表明，这项新技术对于将基因转染到哺乳动物细胞中是有用的。
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