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

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


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cells, Cultured
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / embryology
  • Embryo, Mammalian / metabolism*
  • Female
  • Flow Cytometry
  • Gene Knockout Techniques
  • Humans
  • Immunohistochemistry
  • Integrases / genetics
  • Integrases / metabolism
  • Male
  • Mice, Transgenic
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • RNA, Untranslated / genetics
  • Receptors, Nerve Growth Factor / genetics
  • Receptors, Nerve Growth Factor / metabolism
  • Recombination, Genetic*
  • Sertoli Cells / metabolism*
  • Steroidogenic Factor 1 / genetics
  • Steroidogenic Factor 1 / metabolism


  • Gt(ROSA)26Sor non-coding RNA, mouse
  • NGFR protein, human
  • Nerve Tissue Proteins
  • RNA, Untranslated
  • Receptors, Nerve Growth Factor
  • Steroidogenic Factor 1
  • Cre recombinase
  • Integrases