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Development of a general‐purpose method for cell purification using C re/lox P ‐mediated recombination
Author(s) -
Kuroki Shunsuke,
Akiyoshi Mika,
Ideguchi Ko,
Kitano Satsuki,
Miyachi Hitoshi,
Hirose Michiko,
Mise Nathan,
Abe Kuniya,
Ogura Atsuo,
Tachibana Makoto
Publication year - 2015
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22863
Subject(s) - somatic cell , embryonic stem cell , genetically modified mouse , microbiology and biotechnology , antigen , biology , cell culture , antibody , sertoli cell , embryo , cell , transgene , cell type , chemistry , biochemistry , gene , genetics , spermatogenesis , endocrinology
Summary 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. genesis 53:387–393, 2015. © 2015 Wiley Periodicals, Inc.