PMC:4608092 / 6130-7507 JSONTXT

{"project":"2_test","denotations":[{"id":"26126547-23850618-55294617","span":{"begin":543,"end":545},"obj":"23850618"},{"id":"26126547-18035408-55294618","span":{"begin":546,"end":548},"obj":"18035408"},{"id":"26126547-19423866-55294619","span":{"begin":549,"end":551},"obj":"19423866"}],"text":"Cell Culture and Reagents\nHeLa cells and human embryonic kidney (HEK) 293 cells were maintained in DMEM (Invitrogen, Carlsbad, CA) containing 10% FBS (Japan Bioserum, Hiroshima, Japan). HUVECs (Lonza, Basel, Switzerland) were maintained in endothelial growth medium-2 (Lonza). iPSECs were established and maintained in primate ES medium (ReproCELL, Kanagawa, Japan), supplemented with 500 U/mL of penicillin/streptomycin (Invitrogen) and 4 ng/mL of recombinant human basic fibroblast growth factor (WAKO, Osaka, Japan), as previously reported.14,19,20 For use and establishment of iPS cells, the institutional ethics committees of Kyoto University reviewed and approved the present study protocols. All participants gave written informed consent; for those considered too young to consent, informed consent was given by the parent or guardian.\nRecombinant human IL-1β, TGF-β, VEGF, PDGF-BB, IFN-β, and IFN-γ were obtained from PeproTech (Rocky Hill, NJ). IFN-α was obtained from BioVision (Milpitas, CA). Reagents were dissolved in 0.1% BSA (GIBCO, Carlsbad, CA) at 0 ng/mL.\nFor evaluation of cellular proliferation, HUVECs were seeded at 7.5×104 cells/well in 6-well plates. At 24 hours after seeding, the culture medium including the test reagent was exchanged. The numbers of viable cells were assessed and counted using trypan blue (Nacalai Tesque, Kyoto, Japan) exclusion."}