[Establishment of a high germline competent C57BL/6J ES cell line]

Eleven ES cell lines from C57BL/6J mice were established, with the primary culture of mice embryos as feeder cells and 1 x 10(3) units Leukemia Inhibitory Factor (LIF) in the DMEM (high glucose) medium. The frequency of establishment was 9.6%. Five of these ES cell lines were demonstrated karyotypically normal with diploid composition of over 70%. They shared the characteristics of early embryonic cells: positive for alkaline phosphatase activity and oct4 gene expression. They also showed high potency to differentiate into wide types of cells in vivo. Following injection of the blastocysts, three of them showed abilities to give rise to chimeras and MESPU35 cell line was identified to have high efficiency of colonization into the germline. The clones of MESPU35 cells maintained the germline competence and the mutant clone also retained the high potency to participate into the development of embryos. MESPU35 cells can serve as a valuable vehicle for the production of mutant mice.