JournalofVisualizedExperimentswww.jove.comCopyright©2011JournalofVisualizedExperimentsPage1of2MaterialsListfor:RobustGenerationofHepatocyte-likeCellsfromHumanEmbryonicStemCellPopulationsClaireN.Medine1,BaltasarLucendo-Villarin1,WenliZhou1,ChristopherC.West1,DavidC.Hay11MedicalResearchCouncilCentreforRegenerativeMedicine,UniversityofEdinburghURL:http://www.jove.com/video/2969DOI:doi:10.3791/2969MaterialsMatrigelcoatingplatesandflasks1.Matrigel(10mL,BDBiosciences,UK);storeat-20°C.2.KO-DMEM(500mL,Gibco,Invitrogen,UK);storeat4°C.3.Tissuecultureplates(6well,12well,Corning,UK)4.Tissuecultureflask(25cm2vented,Corning,UK)hESCMaintenance1.Mouseembryonicfibroblastconditionedmedium(MEF-CM)(100mL,R&DSystems,USA);storeat-20°C.2.BSAsolution(50mL,SigmaAldrich,UK);storeat4°C.3.Humanbasicfibroblastgrowthfactor(100μg,Peprotech,USA);storeat-20°C.PassaginghESCswithcollagenase1.ConfluentwellorflaskofhESCs.2.Matrigelcoatedwellsorflasksasappropriate.3.Phospatebufferedsaline(-MgCl2,-CaCl2)(500mL,Gibco,Invitrogen,UK);storeatroomtemperature.4.CollagenaseIV(1g,Gibco,Invitrogen,UK);storeat4°C.5.Mouseembryonicfibroblastconditionedmedium(MEF-CM)(100mL,R&DSystems,USA).6.Humanbasicfibroblastgrowthfactor(100μg,Peprotech,USA).DifferentiationofhESCstohepaticendoderm1.RPMI1640(500mL,Gibco,Invitrogen,UK);storeat4°C.2.B27Supplement(10mL,Gibco,Invitrogen,UK);storeat-20°C.3.ActivinA(2μg,Peprotech,USA);storeat-20°C.4.RecombinantmouseWnt3a(2μg,R&DSystems,USA);storeat-20°C.5.KO-DMEM(500mL,Gibco,Invitrogen,UK);storeat4°C.6.KO-SR(500mL,Gibco,Invitrogen,UK);storeat-20°C.7.Non-essentialaminoacids(100mL,Gibco,Invitrogen,UK);storeat4°C.8.β-Mercapt–thanol(10mL,Gibco,Invitrogen,UK);storeat4°C.9.DMSO(SigmaAldrich,UK);storeatroomtemperature10.LeibovitzL-15culturemedium(500mL,SigmaAldrich,UK);storeat4°C.11.Tryptosephosphatebroth(100mL,SigmaAldrich,UK);storeat4°C.12.F–talbovineserum,heatinactivated(500mL,Gibco,Invitrogen,UK);storeat-20°C.13.Hydrocortisone21-hemisuccinate(100mg,SigmaAldrich,UK...
