media – mouse es cell maintenance media：媒体–小鼠es细胞维持液

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1、SerumBasedmESCmaintenancemediavDMEMUsuallymake4Latatime.Addingredientsinthefollowingorder:IngredientsStocksWorkingvolumesDMEM(GibcoPowderw/oHepes,w/oNaHCo3)Oneliterpacks(Cat#12100-046)4packsNaHCo33.025g/L12.1gCellgrowater(Stirbaron)500ml6bottlesHepes1Mstock25mM(=1000mM/25mM=40,4L/40=100ml)Peni

2、cillin/Streptomycin100X40mlCellgrowater500ml2bottles,bringsolutionup(stirbaroff)to4LbyeyeFilter-sterilizeinhood.Aliquotintoeight500mlbottles.Labelwithcontentsanddate.Glutamine100X1X=Add5mlofGlutaminejustbeforeuse.ORvDMEM-CellGroUsuallyreconstitute500mlatatime.IngredientsStocksWorkingvolumesDME

3、M(MT15-017-CV1X500mlPenicillin/Streptomycin100X5mlGlutamine100X5mlvIMDMUsuallymake4Latatime.Filter-sterilizeinhood.Aliquotintoeight500mlbottles.LabelwithcontentsanddateIngredientsStocksWorkingvolumesIMDM(GibcoPowderw/oHepes,w/oNaHCo3)Oneliterpacks(Cat#12200-069)4packsNaHCo33.025g/L12.1gCellgro

4、water(Stirbaron)500ml6bottlesPenicillin/Streptomycin100X40mlCellgrowater500ml2bottles,bringsolutionup(stirbaroff)to4LbyeyeFilter-sterilizeinhood.Aliquotintoeight500mlbottles.Glutamine100X1X=Add5mlofGlutaminejustbeforeuse.HomemadeIMDM:vAddinabucketof10liters(reservedforcellcultureonly):vPowder1

5、X10litercat#12200-069gibcov8litersofCellgrowatercat#25-055-CVvAdd30.2gramsofNaHCO3(sigmacat#S-5761)vStiratRTforatleast1hrvAddCellgrowaterforupto10litersvStiragainvFiltervAliquot750mlin1literbottleandkeepat4Cfor1-2monthsprotectedfromlight.ORvIMDMUsuallyreconstitute500mlatatime.IngredientsStocks

6、WorkingvolumesIMDM(Cellgro#15-016-CV)1X500mlPenicillin/Streptomycin100X5mlGlutamine100X5mlvDMEM-ES(Serumcontainingmedia)IngredientsStockWorkingvolumes125mlHeat-inactivatedES-QualifiedGemCell100%20%(Tested10-20%)25mlLIF(batchdependent)100%0.5%-5%or10ng/ml0.625-6.25mlMTG1.25gm/literDilute100ulMT

7、Gstockwith900ulDMEM.Thenadd12.6uL/100mlofDMEM-ESmedia15.75uLHomemadeDMEM1X1XMakeupvolumeto125mlvIMDM-ES(Serumcontainingmedia)IngredientsStockWorkingvolumes125mlHeat-inactivatedES-QualifiedGemCell100%20%(Tested10-20%)25mlLIF(batchdepende