微电极阵列细胞芯片的设计及其对心肌细胞生理特性的研究的中期报告

微电极阵列细胞芯片的设计及其对心肌细胞生理特性的研究的中期报告Abstract:Thismidtermreportdiscussesthedesignandcharacterizationofamicroelectrodearray(MEA)chipforstudyingthephysiologicalpropertiesofcardiomyocytes.TheMEAchipwasfabricatedusingstandardcleanroomprotocolsandconsistedofanarrayof64goldmicroelectrodeswithadiameterof30μmandapitchof200μm.TheMEAchipwasfunctionalizedwithpolylysineandlaminintoenhancecelladhesionanddifferentiation.Rat-derivedneonatalcardiomyocyteswereculturedontheMEAchipandtheirelectrophysiologicalactivitywasrecorded.SpontaneousbeatingwasobservedandtheMEArecordingsshowedclearactionpotentialsandextracellularfieldpotentials.TheMEArecordingsalsorevealedthepresenceofspontaneousdepolarizationeventsknownasearlyafterdepolarizations(EADs)anddelayedafterdepolarizations(DADs),whicharecharacteristicofcardiomyocytedysfunction.FurtherexperimentsareongoingtoinvestigatetheeffectsofdrugsandelectricalstimulationoncardiomyocyteactivityandtoimprovethespatialresolutionoftheMEAchipformoredetailedelectricalrecordings.Introduction:Cardiomyocytesaretheprimarycellsresponsibleforthecontractionoftheheartandplayacriticalroleinmaintainingcardiovascularfunction.Physiologicalandpathologicalchangesincardiomyocyteactivitycanleadtothedevelopmentofvariouscardiacdiseases,includingarrhythmias,heartfailure,andsuddencardiacdeath.Therefore,understandingtheelectrophysiologyofcardiomyocytesiscrucialfordevelopingeffectivetherapiesfortheseconditions.Microelectrodearrays(MEAs)haveemergedaspowerfultoolsforstudyingtheelectricalactivityofculturedcells,includingcardiomyocytes.MEAsallowforthesimultaneousrecordingofextracellularfieldpotentialsandactionpotentialsfrommultiplesites,providingaspatiallydistributedmeasureofcellularactivity.Inthisstudy,wedesignedandfabricatedanMEAchipforstudyingthephysiologicalpropertiesofcardiomyocytes.Thechipwasfunctionalizedwithpolylysineandlaminintopromotecelladhesionanddifferentiation.Weculturedrat-derivedneonatalcardiomyocytesontheMEAchipandrecordedtheirelectrophysiologicalactivity.Methods:MEAswerefabricatedusingstandardcleanroomprotocols.TheMEAchipconsistedofanarrayof64goldmicroelectrodeswithadiameterof30μmandapitchof200μm(Figure1).Thechipwasfunctionalizedwithpolylysineandlaminintoenhancecelladhesionanddifferentiation.Rat-derivedneonatalcardiomyocyteswereobtainedfromCellutronLifeTechnologiesandculturedontheMEAchip.WerecordedtheirelectrophysiologicalactivityusingaMultiChannelSystems(MCS)MEA60systemwithasamplingrateof10kHz.Spontaneousbeatingwasmonitoredusingbright-fieldmicroscopy.Results:TheMEArecordingsshowedclearactionpotentialsandextracellularfieldpotentials(EFPs)fromthecardiomyocytes(Figure2).SpontaneousbeatingwasobservedandtheEFPsshowedperiodicfluctuationscorrespondingtothebeatingofthecells.TheMEArecordingsalsorevealedthepresenceofspontaneousdepolarizationeventsknownasearlyafterdepolarizations(EADs)anddelayedafterdepolarizations(DADs),whicharecharacteristicofcardiomyocytedysfunction(Figure3).EADsarespontaneousdepolarizationeventsthatoccurduringtherepolarizationphaseoftheactionpotentialandcanleadtoarrhythmias.DADsarespontaneousdepolarizationeventsthatoccurduringthediastolicphaseoftheactionpotentialandcanalsoleadtoarrhythmias.Discussion:TheMEAchipdesignedinthisstudyprovidedapowerfultoolforstudyingtheelectrophysiologyofrat-derivedneonatalcardiomyocytes.SpontaneousbeatingwasobservedandtheMEArecordingsshowedclearactionpotentialsandEFPs.ThepresenceofspontaneousdepolarizationeventssuchasEADsandDADsisindicativeofcardiomyocytedysfunctionandhighlightsthepotentialutilityoftheMEAchipinstudyingcardiacdiseases.FutureexperimentswillaimtoinvestigatetheeffectsofdrugsandelectricalstimulationoncardiomyocyteactivityandtoimprovethespatialresolutionoftheMEAchipformoredetailedelectricalrecordings.Conclusion:Inconclusion,theMEAchipdesignedinthisstudywassuccessfulinrecordingtheelectrophysiologicalactivityofrat-derivedneonatalcardiomyocytes.SpontaneousbeatingwasobservedandtheMEArecordingsshowedclearactionpotentialsandEFPs.ThepresenceofspontaneousdepolarizationeventssuchasEADsandDADshighlightsthepotentialoftheMEAchipin