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

• Published in: Journal of Reproduction and Development 2012, Vol.58(1), pp.77-83
• Main Authors: KANDA, Akifumi, SOTOMARU, Yusuke, SHIOZAWA, Seiji, HIYAMA, Eiso
• Format: Article
• Language: English
• Subjects: Chimera; Embryonic stem cells; Gene expression; Murine
• ISSN: 0916-8818; 1348-4400
• DOI: 10.1262/jrd.10-178A

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.