多次甘油休克可提高非同步化CHO细胞的磷酸钙转染效率。

Multiple glycerol shocks increase the calcium phosphate transfection of non-synchronized CHO cells.

作者信息

Grosjean Frederic, Bertschinger Martin, Hacker David L, Wurm Florian M

机构信息

Institute of Biochemistry, Chemin des Boveresses 155, 1066, Epalinges, Switzerland.

出版信息

Biotechnol Lett. 2006 Nov;28(22):1827-33. doi: 10.1007/s10529-006-9167-7. Epub 2006 Sep 29.

DOI:10.1007/s10529-006-9167-7

PMID:17009090

Abstract

The exposure of CHO DG44 cells to an osmotic shock, after DNA uptake, results in a cellular volume decrease of approx. 55%. Repetitive osmotic shocks targeted different sub-populations of cells as was demonstrated using two different fluorescent reporter genes. Also the exposure of a calcium phosphate-DNA coprecipitate to high osmolarity in vitro caused the release of the DNA from the precipitate. The results demonstrate the importance of the osmotic shock on the efficient delivery of plasmid DNA to the nucleus of CHO cells following calcium phosphate-mediated transfection.

摘要

在摄取DNA后，将CHO DG44细胞暴露于渗透压休克下，会导致细胞体积大约减少55%。如使用两种不同的荧光报告基因所证明的，重复性渗透压休克针对不同的细胞亚群。此外，体外将磷酸钙-DNA共沉淀物暴露于高渗透压会导致DNA从沉淀物中释放。这些结果证明了渗透压休克在磷酸钙介导的转染后将质粒DNA有效递送至CHO细胞核中的重要性。