Development of a general‐purpose method for cell purification using Cre/loxP‐mediated recombination |
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| Authors: | Shunsuke Kuroki Mika Akiyoshi Ko Ideguchi Satsuki Kitano Hitoshi Miyachi Michiko Hirose Nathan Mise Kuniya Abe Atsuo Ogura Makoto Tachibana |
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| Affiliation: | 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 |
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| 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. genesis 53:387–393, 2015. © 2015 Wiley Periodicals, Inc. |
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| Keywords: | immunological cell purification hCD271 Cre/loxP‐mediated recombination mouse |
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