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Genesis. 2015 Jun;53(6):387-93. doi: 10.1002/dvg.22863. Epub 2015 Jun 11.

Development of a general-purpose method for cell purification using Cre/loxP-mediated recombination.

Author information

1
Department of Enzyme Chemistry, Institute for Enzyme Research, Tokushima University, Tokushima, Japan.
2
Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Sakyo-Ku, Kyoto, Japan.
3
Graduate School of Biostudies, Kyoto University, Sakyo-Ku, Kyoto, Japan.
4
Bioresource Center, RIKEN Tsukuba Institute, Tsukuba, Japan.

Abstract

A mammalian body is composed of more than 200 different types of cells. The purification of a certain cell type from tissues/organs enables a wide variety of studies. One popular cell purification method is immunological isolation, using antibodies against specific cell surface antigens. However, this is not a general-purpose method, since suitable antigens have not been found in certain cell types, including embryonic gonadal somatic cells and Sertoli cells. To address this issue, we established a knock-in mouse line, named R26 KI, designed to express the human cell surface antigen hCD271 through Cre/loxP-mediated recombination. First, we used the R26 Kl mouse line to purify embryonic gonadal somatic cells. Gonadal somatic cells were purified from the R26 KI; Nr5a1-Cre-transgenic (tg) embryos almost equally as efficiently as from Nr5a1-hCD271-tg embryos. Second, we used the R26 KI mouse line to purify Sertoli cells successfully from R26 KI; Amh-Cre-tg testes. In summary, we propose that the R26 KI mouse line is a powerful tool for the purification of various cell types.

KEYWORDS:

Cre/loxP-mediated recombination; hCD271; immunological cell purification; mouse

PMID:
26012873
DOI:
10.1002/dvg.22863
[Indexed for MEDLINE]

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