201B6 (International Stem Cell Registry)

Growth conditions: Generate and maintain iPS cells in primate ES medium (ReproCELL, Japan) supplemented with recombinant human basic fibroblast growth factor (bFGF, 4 ng/ml). Seed iPS cells on Matrigel-coated plates in mouse embryonic fibroblast (MEF)-conditioned primate ES cell medium, supplemented with bFGF (4 ng/ml). Change the medium daily. For preparation of MEF-conditioned medium: Plate MEFs derived from embryonic day 13.5 embryo pool of ICR mice at 1*10^6 cells per 100 mm dish and incubate overnight. Then, wash the cells once with PBS and culture in 10 ml primate ES cell medium. After a 24 hr incubation, collect the supernatant of the MEF culture, filter through a 0.22 mm pore-size filter and store.