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PluripotentStemCellsandReprogrammingContentsContentsWhatarestemcells?Blastocyst


Implantation


Embryonicdevelopment


Postnataldevelopment


AdultContentsEmbryonicstemcells:Embryonicstemcells(ESCs)JamesThomson,Ph.D.SirMartinJohnEvansisaBritishscientistwho,withMatthewKaufman,wasthefirsttoculturemiceembryonicstemcellsandcultivatetheminalaboratoryin1981.Heisalsoknown,alongwithMarioCapecchiandOliverSmithies,forhisworkinthedevelopmentoftheknockoutmouseandtherelatedtechnologyofgenetargeting,amethodofusingembryonicstemcellstocreatespecificgenemodificationsinmice.In2007,thethreesharedtheNobelPrizeinPhysiologyorMedicineinrecognitionoftheirdiscoveryandcontributiontotheeffortstodevelopnewtreatmentsforillnessesinhumans.EmbryonicGermCells(EGCs)JohnD.Gearhart,Ph.D.

StemCellBiologyProgram
JohnsHopkinesUniversitySchoolofMedicineMouseearlydevelopmentMouseESCderivation◆Feedercellsandserum
Feedercellsprovideleukemiainhibitoryfactor(LIF)
Serumprovidesbonemorphogeneticproteins(BMPs)
◆Feeder-andserum-free
LIFandBMP
◆LIF-andBMP-free(2i)
Thefibroblastgrowthfactor/extracellularsignal-relatedkinase(Fgf/Erk1/2)inhibitor
Glycogensynthasekinase3(GSK3)inhibitor
Sun,etal.HumanearlydevelopmentInvitrodevelopmentofIVF体外受精frozenembryosHumanearlyembryosMichaelAmitetal,JAnat.200:225,2002DerivationofthreenewChinesehumanEScelllines.(A)MorphologyofasurplushumanblastocystfromtheIVFclinics;(B)PrimaryICMoutgrowth(Passage0);(C)Typicalundifferentiatedcoloniespassagedbymechanicalsplitting(Passage5);(D)HighermagnificationofahumanESCcolonypassagedbymechanicalsplitting(Passage9).Lietal.(2010)
InvitroCell&Dev.Biol.–Animal46:186-191.CultureofhumanESCsMechanicalpassageprocedureofhumanEScells.

Typicalundifferentiatedcolonieswereseparatedfromfeederlayersusingglassneedles.Thencolonieswerecuthorizontally,andthenverticallytoformuniformclumpsandtransferredontonewlypreparedfeederlayers.Feeder-freeculturehEScellsFeedercellsderivedfromhumantissue:NatureBiotech
2006DerivationofhESCsindefinedconditions2006Establishmentofhum