Hepatocyte Differentiation from Human iPSCs

Human iPSCs CGT-RCiB-10 (Cell & Gene Therapy Catapult) were maintained on Vitronectin XF (STEMCELL Technologies) coated Corning Costar TC‐treated six‐well plates (Sigma Aldrich) in Essential 8 Medium (Thermo Fisher Scientific) and passaged every 4 days using Gentle Cell Dissociation Reagent (STEMCELL Technologies).
Hepatocyte differentiation was carried out as previously described^{58 (link),59}. Silencing of human RBFOX2 was performed by transfecting 100 nM smart pool to RBFOX2 or a mock control (Horizon) with RNAimax reagent (Invitrogen) in OptiMEM (Invitrogen).

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Paterson H.A., Yu S., Artigas N., Prado M.A., Haberman N., Wang Y.F., Jobbins A.M., Pahita E., Mokochinski J., Hall Z., Guerin M., Paulo J.A., Ng S.S., Villarroya F., Rashid S.T., Le Goff W., Lenhard B., Cebola I., Finley D., Gygi S.P., Sibley C.R, & Vernia S. (2022). Liver RBFOX2 regulates cholesterol homeostasis via Scarb1 alternative splicing in mice. Nature Metabolism, 4(12), 1812-1829.

Publication 2022

Vitronectin XF Essential 8 Medium Gentle Cell Dissociation Reagent RNAimax reagent OptiMEM

Cell Cell therapy Gene Gene therapy Hepatocyte Human Ipscs Smart 100 Stemcell Vitronectin

Corresponding Organization :

Other organizations : MRC London Institute of Medical Sciences, Harvard University, Hammersmith Hospital, Imperial College London, Fondation pour l’innovation en Cadiométabolisme et Nutrition, Inserm, Sorbonne Université, Institute for Research in Biomedicine, Universitat de Barcelona, University of Edinburgh

Top 5 similar protocols

Variable analysis

independent variables

Silencing of human RBFOX2 by transfecting 100 nM smart pool to RBFOX2

dependent variables

Hepatocyte differentiation

control variables

Human iPSCs CGT-RCiB-10 maintained on Vitronectin XF coated Corning Costar TC-treated six-well plates in Essential 8 Medium and passaged every 4 days using Gentle Cell Dissociation Reagent
Mock control (Horizon) transfected with RNAimax reagent in OptiMEM

controls

Positive control: Hepatocyte differentiation as previously described in references 58 and 59
Negative control: Mock control (Horizon) transfected with RNAimax reagent in OptiMEM

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