DifferentiateEScellsintocysticembryoidbodies Day-1:PassEScellsatnormaldensityongelatinizedplatetofreethecultureofcontaminationfibroblastcells. ______________ Day1:Trypsinizedthecellsasfornormalpassaginguntilthecoloniesliftoff.Trytokeepthelooselyconnectedclumpsofcellstogetherbygentlehandling.Thendirectlyplatethecells1:3intobacterialgradePetridishesinLIFfreeEScellmedium. Day3:Aspiratethemediumcarefully.Avoidsuckinguptoomanyoftheaggregates.Thenaddnewmedium. Day7:Changethemedium Day9:Changethemedium Day11:Changethemedium Day13:Changethemedium Day15:Bythisdayahighpercentageoftheembryoidbodieswillhavedevelopedfluidfilledcavities.Thestructuresarecalledcysticembryoidbodies