Establishment of ES Cells from Inbred Strain Mice by Dual Inhibition (2i)

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  • KANDA Akifumi
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima 739-8511, Japan
  • SOTOMARU Yusuke
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima 739-8511, Japan
  • SHIOZAWA Seiji
    Central Institute for Experimental Animals, Kanagawa 210-0821, Japan Department of Physiology, Keio University School of Medicine, Tokyo 160-8582, Japan
  • HIYAMA Eiso
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima 739-8511, Japan

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Abstract

A number of mouse ES cells from inbred strains have been established to date, but efficiency varies across the different strains. The 129 strain mouse is efficient to establish, whereas C57BL/6 and BALB/c strains are not. It is possible that their genetic backgrounds account for the difference in their ability to establish ES cell lines. In this study, we attempted to establish C57BL/6J and BALB/c Cr ES cells by dual inhibition (2i) using two inhibitors (PD0325901 and CHIR99021) of extracellular signal regulated-kinase (ERK) and glycogen synthase kinase-3 (GSK-3), which promote ES cell differentiation. The results revealed that the establishment efficiencies of C57BL/6J and BALB/c Cr ES cells were remarkably increased by 2i. These ES cells stably expressed pluripotent markers and generated high-contribution chimeras with germline transmission. Furthermore, we generated germline chimeras from C57BL/6J ES cells through the method of gene modification. These findings indicate that 2i is a powerful tool for establishing C57BL/6J and BALB/c Cr ES cells with the ability to generate germline chimeras.

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