用大鼠原代心肌细胞构建工程化心肌组织的研究

《用大鼠原代心肌细胞构建工程化心肌组织的研究》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、用大鼠原代心肌细胞构建工程化心肌组织的研究作者：常海霞，郑强荪,刘兴光【摘要】目的探索用大鼠原代心肌细胞构建工程化心肌组织的方法。方法用胶原、Matrigel、2倍DMEM分别与含%EDTA的%胰酶制备的乳鼠原代心肌细胞和%胰酶制备的原代心肌细胞混合，注入环形槽内形成心肌条带，一周后行力学拉伸继续培养一周。常规HE染色等对组织工程化心肌组织，即心肌条带进行评价。结果心肌条带出现自发性跳动，HE染色显示与正常成年大鼠心肌组织类似，电镜结果显示条带中的心肌细胞含有丰富的肌原纤维,肌小节结构和Z带清晰可见。用含%EDTA的%胰酶制备的原代乳鼠心肌细胞构建的心肌条带，拉伸后第4天形成一致性跳动。

2、免疫组化检测提示肌钙蛋白阳性细胞数多于对照组。电镜观察显示，条带的心肌细胞中肌原纤维较对照组明显增多。结论这一方法可构建工程化心肌组织，心肌细胞制备方法的改进以及液态Ⅰ型胶原浓度的精确测定，可显著提高构建心肌条带的成功率及条带的生理功能，使其更接近正常心肌组织。【关键词】原代心肌细胞;液态Ⅰ型胶原;工程化心肌组织基金项目：全军医学科研“十五”计划项目自主ExperimentalStudyofEngineeredHeartTissueUsingNeonatalRatPrimaryCardiomyocytesAbstract:OBJECTIVEToconstructengineeredhea

3、rttissue(EHT)usingneonatalratprimaryLiquidcollagentypeI,Matrigeland×DMEMweremixedwithneonatalratcardiacmyocytesisolatedusing%trypsinincluding%EDTA,or%trypsinonlyasacontrol.Themixturewasthenpippettedintocircularmoldstoconstructcardiacring.dayslater,thecardiacringwassubjectedto10%staticstretchforan

4、otherdays.microscopyandtransmissionelectronmicroscopy,routineHEstainingandimmunohistochemicalstainingwereusedtoanalyzetheSpontaneousbeatingwasseenintheEHTs.HEstainingrevealedthatcardiacmyocyteswithintheEHTsresembledthatofthenativeadultcardiactissueinmorphology.Transmissionelectronmicroscopyreveal

5、edthatthecardiacmyocyteswithintheEHTscontainedabundantmyofibrils,clearlydefinedsarcomeresandZlines.TheEHTsconstructedusingthecardiacmyocytesisolatedby%trypsinsupplementedwith%EDTAfinallycametosynchronousbeatingatthefourthdayofstretching.Histologicalandimmunohistochemicalanalysisalsoshowedmoreposi

6、tivecardiacmyocytesandmoremyofibrilspercellthaninthecontrolThemethoddescribedinthisstudyishelpfulinconstructingtissueengineeredhearttissue.TheimprovedtechniqueforcardiacmyocyteisolationandaccuratequantificationofliquidcollagentypeIarebeneficialtoEHTsconstructionandtheEHTsarelikelytosharemorestruc

7、turalandfunctionalsimilaritieswiththatofsimilartothenativehearttissue.Keywords：Primarycardiacmyocyte;LiquidcollagentypeI;Engineeredhearttissue(EHT)本实验旨在对在体外构建和重现心脏组织的方法进行探索，寻找能够构建类似正常心肌组织的方法，这一方法XX年德国的科学家Zimmermann等人的研