Feeder-free Culture of Human iPSCs and ESCs

The experiments using human iPSCs and ESCs were approved by the Ethics Committee of the Department of Medicine and Graduated School of Medicine, Kyoto University. Informed consent was obtained from all donors from whom iPSCs were derived. The iPSC line 585A1^{35 (link)} was cultured in Essential 8 medium (Thermo Fisher Scientific). The ESC line KhES-3^{36 (link)} and iPSC line Ff-I01^{37 (link)} were cultured in StemFit AK02N medium (Reprocell). The cells were maintained under feeder-free conditions, passaged every 3–4 days using 0.5 mM EDTA/PBS (Thermo Fisher Scientific) and gentle pipetting, and routinely examined for mycoplasma contamination.

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Ito R., Kimura A., Hirose Y., Hatano Y., Mima A., Mae S.I., Keidai Y., Nakamura T., Fujikura J., Nishi Y., Ohta A., Toyoda T., Inagaki N, & Osafune K. (2023). Elucidation of HHEX in pancreatic endoderm differentiation using a human iPSC differentiation model. Scientific Reports, 13, 8659.

Publication 2023

Essential 8 medium StemFit AK02N medium EDTA/PBS

Cells Cultured medium Donors Edta Escs Human Ipsc Mycoplasma

Corresponding Organization :

Other organizations : Kyoto University

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Variable analysis

independent variables

Cell lines used: iPSC line 585A1, ESC line KhES-3, and iPSC line Ff-I01

dependent variables

Outcomes of culturing the cell lines in Essential 8 medium and StemFit AK02N medium

control variables

Feeder-free culture conditions
Passaging the cells every 3-4 days using 0.5 mM EDTA/PBS and gentle pipetting
Routine examination for mycoplasma contamination

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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