A microwell array system for stem cell culture

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1、ARTICLEINPRESSBiomaterials29(2008)752–763www.elsevier.com/locate/biomaterialsAmicrowellarraysystemforstemcellculturea,bca,dHannes-ChristianMoeller,MatthewK.Mian,ShamitShrivastava,a,ca,c,BongGeunChung,AliKhademhosseiniaDepartmentofMedicine,CenterforBiomedicalEngineering,BrighamandWomensHospital,H

2、arvardMedicalSchool,Boston,MA02115,USAbE´coleSupe´rieuredeBiotechnologiedeStrasbourg,F-67412Illkirch-Cedex,FrancecHarvard-MITDivisionofHealthSciencesandTechnology,MassachusettsInstituteofTechnology,Cambridge,MA02139,USAdDepartmentofMechanicalEngineering,CenterforNanotechnology,IndianInstituteofTe

3、chnology,Guwahati,IndiaReceived1August2007;accepted17October2007Availableonline14November2007AbstractDirectedembryonicstem(ES)celldifferentiationisapotentiallypowerfulapproachforgeneratingarenewablesourceofcellsforregenerativemedicine.TypicalinvitroEScelldifferentiationprotocolsinvolvetheformatio

4、nofEScellaggregateintermediatescalledembryoidbodies(EBs).Recently,wedemonstratedtheuseofpoly(ethyleneglycol)(PEG)microwellsastemplatesfordirectingtheformationoftheseaggregates,offeringcontroloverparameterssuchassize,shape,andhomogeneity.Despitethesepromisingresults,thepreviouslydevelopedtechnolog

5、ywaslimitedasitwasdifficulttoreproduciblyobtainculturesofhomogeneousEBswithhighefficiencyandretrievability.Inthisstudy,weimprovetheplatformbyoptimizinganumberoffeatures:materialcompositionofthemicrowells,cellseedingprocedures,andaggregateretrievalmethods.Adoptingthesemodifications,wedemonstrateanimp

6、roveddegreeofhomogeneityoftheresultingaggregatepopulationsandestablisharobustprotocolforelicitinghighEBformationefficiencies.TheoptimizedmicrowellarraysystemisapotentiallyversatiletoolforEScelldifferentiationstudiesandhigh-throughputstemcellexperimentation.r2007ElsevierLtd.Allrightsreserved.Keywor

7、ds:Microwells;Poly(ethyleneglycol)(PEG);Embryoidbodies(EBs)1.Introductionawidespectrumofcelltypes[13–16].EBsareusuallyformedusingthehangingdropmethod[16,17]orinEmbryonicstem(ES)cellsholdtherapeuticpotentialassuspension