title: Development of a general-purpose method for cell purification using Cre/loxP-mediated recombination.
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.
authors.affiliation: Department of Enzyme Chemistry, Institute for Enzyme Research, Tokushima University, Tokushima, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Kuroki
authors.affiliation: Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Sakyo-Ku, Kyoto, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Akiyoshi
authors.affiliation: Department of Enzyme Chemistry, Institute for Enzyme Research, Tokushima University, Tokushima, Japan.|Graduate School of Biostudies, Kyoto University, Sakyo-Ku, Kyoto, Japan.
authors.forename: Ko
authors.initials: K
authors.lastname: Ideguchi
authors.affiliation: Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Sakyo-Ku, Kyoto, Japan.
authors.forename: Satsuki
authors.initials: S
authors.lastname: Kitano
authors.affiliation: Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Sakyo-Ku, Kyoto, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Miyachi
authors.affiliation: Bioresource Center, RIKEN Tsukuba Institute, Tsukuba, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Hirose
authors.affiliation: Bioresource Center, RIKEN Tsukuba Institute, Tsukuba, Japan.
authors.forename: Nathan
authors.initials: N
authors.lastname: Mise
authors.affiliation: Bioresource Center, RIKEN Tsukuba Institute, Tsukuba, Japan.
authors.forename: Kuniya
authors.initials: K
authors.lastname: Abe
authors.affiliation: Bioresource Center, RIKEN Tsukuba Institute, Tsukuba, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.affiliation: Department of Enzyme Chemistry, Institute for Enzyme Research, Tokushima University, Tokushima, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Tachibana
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D055786:Gene Knockout Techniques; D006801:Humans; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008297:Male; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012708:Sertoli Cells / Q000378:metabolism*; D054339:Steroidogenic Factor 1 / Q000235:genetics / Q000378:metabolism
chemical_list: C110853:Gt(ROSA)26Sor non-coding RNA, mouse; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D022661:RNA, Untranslated; D017475:Receptors, Nerve Growth Factor; D054339:Steroidogenic Factor 1; C045073:Cre recombinase; D019426:Integrases
keywords: Cre/loxP-mediated recombination; hCD271; immunological cell purification; mouse