USING GROWTH FACTOR ARRAYS AND MICROPATTERNED CO-CULTURES TO INDUCE HEPATIC DIFFERENTIATION OF EMBRYONIC STEM CELLS

《USING GROWTH FACTOR ARRAYS AND MICROPATTERNED CO-CULTURES TO INDUCE HEPATIC DIFFERENTIATION OF EMBRYONIC STEM CELLS》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。

1、Biomaterials31(2010)9221e9231ContentslistsavailableatScienceDirectBiomaterialsjournalhomepage:www.elsevier.com/locate/biomaterialsUsinggrowthfactorarraysandmicropatternedco-culturestoinducehepaticdifferentiationofembryonicstemcellsa,baabca,*NazgulTuleuova

2、,JiYounLee,JenniferLee,ErlanRamanculov,MarkA.Zern,AlexanderRevzinaDepartmentofBiomedicalEngineering,UniversityofCalifornia,Davis,451HealthSciencesDrive,#2519,Davis,CA95616,USAbNationalCenterforBiotechnology,Astana,KazakhstancDepartmentofMedicine,Transplan

3、tResearchInstitute,UniversityofCalifornia,Davis,USAarticleinfoabstractArticlehistory:ThesuccessindrivingembryonicstemcellstowardshepaticlineagehasbeenconfoundedbytheReceived4August2010complexityandcostofdifferentiationprotocolsthatemploylargequantitiesofe

4、xpensivegrowthfactorsAccepted23August2010(GFs).InsteadofsupplementingculturemediawithsolubleGFs,weinvestigatedcultivationandAvailableonline15September2010differentiationofmouseembryonicstemcells(mESCs)onprintedarraysofGFs.Hepatocytegrowthfactor(HGF),basic

5、fibroblastgrowthfactor(bFGF)andbonemorphogeneticprotein(BMP4)weremixedKeywords:insolutionwithfibronectinandcollagen(I)andthenprintedontosilane-modifiedglassslidestoformGrowthfactorarrays500mmdiameterproteinspots.mESCswereculturedontopofGFspotsforupto12daysan

6、danalyzedMicropatternedco-culturesbyRT-PCRandimmunostainingatdifferenttimepoints.ThestemcellsresidingonHGF-containingHepaticdifferentiationEmbryonicstemcellscombinationsofGFsexhibitedrequisitefeaturesofhepaticdifferentiationincludingpronouncedlossinplurip

7、otency(Oct4),transient(upanddown)expressionofendoderm(Sox17)andupregulationofearlyhepaticmarkersealbuminandalpha-fetoprotein.ThehepaticdifferentiationwasenhancedfurtherbyaddinghepaticstellatecellstosurfacesthatalreadycontainedmESCsonGFspots.Acombinationof

8、co-culturewithnon-parenchymallivercellsandtheoptimalGFstimulationwasfoundtoinduceendodermandhepaticphenotypeearlierandtoamuchgreaterextentthantheGFarraysormicropatternedco-culturesusedindividually.Whilethispaperinve