FIAU用于心脏报告基因显像的实验分析

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1、--华中科技大学硕士学位论文结论用Iodogen固相氧化法和Sep-PakC-18反相色谱柱纯化可以快速制备符合实验要求的125I-FIAU，并且所制备的125I-FIAU具有较好的生物分布特性。应用改良的方法可以更简单的培养出符合大多数实验要求的心肌细胞。用腺病毒重组体做载体，可以将HSVI-tk基因成功转导入心肌细胞，并且对125I-FIAU有较高的摄取率。因此，用腺病毒重组体作为载体，HSV1-tk作为标志性基因，125I-FIAU作为标志性底物有望进行心脏报告基因显像。关键词FAU；同位素标记；药代动力学；报告基因；心肌5-----华中科技大学硕士学位论文ExperimentalStu

2、dyofCardiacReporterGeneImagingwithFIAUAbstractObjectiveTheaimofthisstudywastoevaluatethepossibilityofHSV1-tkwhichwastransducedbyanadenoviralvector,andradiolabledFIAUasamarkergene/markersubstrateforimagingreportergeneexpressionintheheart.Inthisstudy,weradiolabeled125I-FIAU,culturedmyocardialcells,con

3、structedrecombinantadenoviruscarryingHSV1-tkgene.Thesusceptibilitytothevector,125I-FIAUuptakeratiosandtherelationstoMOIsandtimeswereevaluatedthroughaseriesoftests.Methods1.FAUwaslabeledwith125I-NaIandthebiodistributionof125I-FIAUinthenormalmicewastoevaluate.FAUwasradiolabeledwith125I-NaIbyIodogeniod

4、ization.TheproductwaspurifiedthroughSep-PakC-18reversephasechromatographcolumn.-----RadiochemicalpurityandthestabilityinvitroweremeasuredbyTLC-SG.Westudiedthebiodistributionof125I-FIAUinthenormalmice,andthedistributionsincardiacmuscleandbloodshouldbeemphasized.1.ToculturenormalSDneonatalratmyocardia

5、lcells.Toculturepurifiedneonatal6-----华中科技大学硕士学位论文ratmyocardialcellsbythesimplifiedandmodifiedmethods.TheneonatalrathearttissuewasdigestedonceincollagenasetypeⅡincludingBSA.Thepurifiedcadiocyteswereobtainedbydifferentialadhesionafter1hour.Allofthesestepswerecarriedoutin25-37℃.Weevaluatedthesurvivalr

6、atethroughstainingthecellwithtypsinonthedayof5th,10th,15th,20th,25th,30th,andthenobservedcellformundertheinvertedbiologicalmicroscopanddetectedthepurityofthecadiocyteswhichwereculturedfor48hoursbythemethodofimmunohistochemistry.3.ToconstructtheAd-CMV-HSV1-tkrecombinant.ThecompoundsofAd-CMV-HSV1-tkwe

7、rerecombinatedbyusingHSV1-tkasareportergene,CMVasapromoterandadenoviralasavector.ThispartoftheresearchwascollaboratedwithVectorGeneCompanyLTD.4.StudyviabilityofthemyocardicalcellsafterAd5-tkinfectiona