iPSCs (lines 15-0001 and 19) were cultured [as previously described (18 (link))] on Geltrex-coated (ThermoFisher) cell culture plates (Greiner) using Essential™ 8 (E8) medium (Gibco, ThermoFisher) supplemented with E8 supplement (50X, Gibco) and 0.5% (v/v) antibiotic-antimycotic (Gibco). Medium was refreshed daily. Upon 80% confluency, iPSCs were washed using PBS and subsequently split into colonies using PBS-EDTA (0.5 mM, ThermoFisher) for 4 to 6 min at room temperature. Cells were split in a ratio of 1:6—1:8 twice per week.
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iPSC Maintenance and Passaging Protocol
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Corresponding Organization :
Other organizations : Royal Netherlands Academy of Arts and Sciences, University Medical Center Utrecht, Roche (Switzerland), Hubrecht Institute for Developmental Biology and Stem Cell Research, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, University Medical Center, Radboud University Nijmegen, RWTH Aachen University, Utrecht University
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Variable analysis
independent variables
- Cell culture substrate (Geltrex-coated cell culture plates)
- Cell culture medium (Essential 8 medium with supplements)
- IPSC growth and proliferation (measured by confluency)
- Antibiotic-antimycotic supplement concentration (0.5% v/v)
- Cell splitting ratio (1:6 - 1:8)
- Cell passage frequency (twice per week)
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