All hiPSC lines used in this study were generated using peripheral blood mononuclear cells. Reprogramming and characterization were performed as previously described49 (link),50 (link). The cells were maintained in vitronectin-coated dishes (Thermo Fisher Scientific, Waltham, MA, USA), and the media were changed daily with fresh Essential 8 (E8) medium (Thermo Fisher Scientific).
Vitronectin coated dishes
Manufactured by Thermo Fisher Scientific
Sourced in United States
Vitronectin-coated dishes are a type of cell culture dish designed to mimic the natural extracellular matrix environment. The dishes are coated with the protein vitronectin, which is a component of the extracellular matrix and promotes cell adhesion and growth.
Automatically generated - may contain errors
2 protocols using vitronectin coated dishes
1
Generation and Maintenance of hiPSC Lines
2
Expansion of Naive Human Pluripotent Stem Cells
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All hPSC experiments were approved by the UK Stem Cell Bank Steering Committee and comply with the regulations of the UK Code of Practice for the Use of Human Stem Cell Lines. Conventional SHEF6 (International Stem Cell et al., 2007 (link)), RUES2, and RUES-GLR (obtained with kind permission from the A.H. Brivanlou laboratory, Rockefeller University) were cultured on vitronectin-coated dishes (10 μg/mL; Thermo Fisher Scientific) in E8 medium (Thermo Fisher Scientific) under hypoxic conditions (37°C, 5% CO2, 5% O2). Cells were routinely passaged in clumps using 50 mM EDTA.
Naive cells were maintained in PXGL (1 μM PD0325901, 2 μM XAV939, 2 μM Gö6983, and 10 ng/mL human LIF in N2B27) on mitomycin C (Sigma) inactivated MEFs (Cambridge Stem Cell Institute) and passaged with Accutase (Thermo Fisher Scientific). Y-27632 (10 μM) was supplemented during the first 24 h after passage. PD0325901, basic FGF, XAV939, Gö6983, and human LIF were obtained from the Cambridge Stem Cell Institute.
Naive cells were maintained in PXGL (1 μM PD0325901, 2 μM XAV939, 2 μM Gö6983, and 10 ng/mL human LIF in N2B27) on mitomycin C (Sigma) inactivated MEFs (Cambridge Stem Cell Institute) and passaged with Accutase (Thermo Fisher Scientific). Y-27632 (10 μM) was supplemented during the first 24 h after passage. PD0325901, basic FGF, XAV939, Gö6983, and human LIF were obtained from the Cambridge Stem Cell Institute.
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