Effect of Substrate Stiffness on Early Mouse Embryo Development 英文参考文献.docVIP

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Effect of Substrate Stiffness on Early Mouse Embryo Development 英文参考文献.doc

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Effect of Substrate Stiffness on Early Mouse Embryo Development 英文参考文献

EffectofSubstrateStiffnessonEarlyMouseEmbryo Development KevinS.Kolahi1,AnnemarieDonjacour1,XiaoweiLiu1,WingkaLin1,RhodelK.Simbulan1,EnrricoBloise1, EminMaltepe2,PaoloRinaudo1* 1Department of Obstetrics Gynecology and Reproductive Sciences, University of California San Francisco, San Francisco, California, United States of America, 2DepartmentofPediatrics,UniversityofCaliforniaSanFrancisco,SanFrancisco,California,UnitedStatesofAmerica Abstract Itisbecomingincreasinglyclearthatcellsareremarkablysensitivetothebiophysicalcuesoftheirmicroenvironmentand thatthesecuesplayasignificantroleininfluencingtheirbehaviors.Inthisstudy,weinvestigatedwhethertheearlypre- implantation embryo is sensitive to mechanical cues, i.e. the elasticity of the culture environment. To test this, we have developedanewembryoculturesystemwherethemechanicalpropertiesoftheembryonicenvironmentcanbeprecisely defined.Thecontemporarystandardenvironmentforembryocultureisthepolystyrenepetridish(PD),whichhasastiffness (1GPa) that is six orders of magnitude greater than the uterine epithelium (1kPa). To approximate more closely the mechanical aspects of the in vivo uterine environment we used polydimethyl-siloxane (PDMS) or fabricated 3D type I collagen gels (1kPa stiffness, Col-1k group). Mouse embryo development on alternate substrates wascompared to that seenonthepetridish;percentdevelopment,hatchingfrequency,andcellnumberwereobserved.Ourresultsindicated thatembryosaresensitivetothemechanicalenvironmentonwhichtheyarecultured.EmbryosculturedonCol-1kshowed asignificantlygreater frequencyof developmentto2-cell (68615%vs.59618%),blastocyst (6469.1% vs.50618%)and hatching blastocyst stages (54625% vs. 21616%) and an increase in the number of trophectodermal cell (TE,65613 vs. 49612cells)comparedtocontrolembryosculturedinPD(mean6S.D.;p,.01).EmbryosculturedonCol-1kandPDwere transferred to recipient females and observed on embryonic day 12.5. Both groups had the same number of fetuses, howevertheplacentasoft