Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6162516 | Kidney International | 2013 | 6 Pages |
Abstract
Biomedical research often requires primary cultures of specific cell types, which are challenging to obtain at high purity in a reproducible manner. Here we engineered the murine Rosa26 locus by introducing the diphtheria toxin receptor flanked by loxP sites. The resultant strain was nicknamed the Terminator mouse. This approach results in diphtheria toxin-receptor expression in all non-Cre expressing cell types, making these cells susceptible to diphtheria toxin exposure. In primary cultures of kidney cells derived from the Terminator mouse, over 99.99% of cells were dead within 72Â h of diphtheria toxin treatment. After crossing the Terminator with the podocin-Cre (podocyte specific) mouse or the Ggt-Cre (proximal tubule specific) mouse, diphtheria toxin treatment killed non-Cre expressing cells but spared podocytes and proximal tubule cells, respectively, enriching the primary cultures to over 99% purity, based on both western blotting and immunostaining of marker proteins. Thus, the Terminator mouse can be a useful tool to selectively and reproducibly obtain even low-abundant cell types at high quantity and purity.
Related Topics
Health Sciences
Medicine and Dentistry
Nephrology
Authors
Jian-Kan Guo, Hongmei Shi, Farrukh Koraishy, Arnaud Marlier, Zhaowei Ding, Alan Shan, Lloyd G. Cantley,