title: IPC, 2D, no SAG, 17K6 iPSC derived
organism: Homo sapiens
description: FOXD1 reporter iPSCs (17K6) were generated by gene targeting of iPSCs (201B7) and used in this study. hiPSCs were first treated using an enzymatic method with a 1:1 mixture of TrypLE Select Enzyme and 0.5 mM EDTA/PBS for 5 min and washed with PBS(-). The cells were then detached using a cell scraper, dissociated into single cells by gentle pipetting and seeded onto 24-well plates (CORNING) at a density of 1.3*10^4 cells/cm^2 with 0.5 mL of Stem Fit AK02N supplemented with 10 uM Y-27632 (day -1) and 2.4 uL/mL iMatrix-511 (Nippi). After 24 h (day 0), the cells were cultured with serum-free differentiation medium consisting of DMEM/F12 Glutamax medium (Thermo Fisher Scientific), B27 supplement minus vitamin A (Thermo Fisher Scientific), and 0.5X Penicillin/streptomycin (hereafter called basal medium) supplemented with 5 uM CHIR99021 (Wako), 10 nM RA (SIGMA), 1 ng/mL BMP4 (Peprotech), and 100 ng/mL bFGF (Wako). After another 24 h (day 1), the cells were cultured with 5 uM CHIR99021, 100 ng/mL bFGF, and 1 ng/mL BMP7 (R&D Systems). After a third 24 h (day 2), the medium was switched to basal medium containing 5 uM CHIR99021, 1 ng/mL BMP7, and 10 uM A83-01 (Wako). After a fourth 24 h (day 3), the medium was changed to basal medium containing 5 uM CHIR99021, 30 ng/mL bFGF, 10 ng/mL activin A (R&D Systems), and 30 yM Y-27632 (Wako). After a fifth 24 h (day 4), the cells were washed with PBS (-), treated with Accumax (Innovative Cell Technologies, Inc.), dissociated into single cells by gentle pipetting. On day 4 of PPS induction, the cells were seeded on 24-well plates (Corning) at a density of 2.0*10^5 cells/well with basal medium containing 5 uM CHIR99021, 30 ng/mL bFGF, 10 ng/mL activin A, 30 uM Y-27632 and iMatrix, and the cells were cultured for 48 h. On day 6, the cells were incubated with AK02N minus bFGF medium (Ajinomoto) containing 1 uM CHIR99021, 0.1 uM RA and 10 uM SB431542 (Nakalai tesque) for 120 h.
attributes.tag: sample_name
attributes.value: IP0208
attributes.tag: age
attributes.value: not applicable
attributes.tag: biomaterial_provider
attributes.value: Kenji Osafune
attributes.tag: cell_type
attributes.value: hiPS cell derived IPCs, 2D
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: sex
attributes.value: not applicable
attributes.tag: tissue
attributes.value: not applicable
attributes.tag: collection_date
attributes.value: missing
attributes.tag: geo_loc_name
attributes.value: missing
attributes.tag: BioSampleModel
attributes.value: Human