Cardiomyocytes: Methods and Protocols

221 Pages·2015·4.169 MB·English

by  Gary R. Skuse, Maureen C. Ferran (eds.)

Methods in
Molecular Biology 1299
Gary R. Skuse
Maureen C. Ferran Editors
Cardio-
myocytes
Methods and Protocols
M M B
ETHODS IN OLECULAR IOLOGY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire,AL109AB,UK
For further volumes:
http://www.springer.com/series/7651

Cardiomyocytes
Methods and Protocols
Edited by
Gary R. Skuse and Maureen C. Ferran
Rochester Institute of Technology, Rochester, NY, USA
Editors
GaryR.Skuse MaureenC.Ferran
RochesterInstituteofTechnology RochesterInstituteofTechnology
Rochester,NY,USA Rochester,NY,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-4939-2571-1 ISBN978-1-4939-2572-8 (eBook)
DOI10.1007/978-1-4939-2572-8
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SpringerNewYorkHeidelbergDordrechtLondon
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CoverIllustration: CytivaTMPlusCardiomyocytes:ThedeconvolvedImagewasacquiredonDeltaVisionOMXby
AngelaWilliams(GEHealthcare).After14daysofgrowththecellswerestainedforcardiacTroponinI(Red)and
a-Actinin(Green)
Printedonacid-freepaper
HumanaPressisabrandofSpringer
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Preface
Heartdiseaseisresponsibleforuntoldglobalmorbidityandmortality.Traditionalmedical
approaches to the treatment of heart disease often strive to ameliorate damage and to
prevent future damage, but they cannot reverse what has already happened. This is
especially apparent in the most extreme instances where heart transplantation is used to
replacetheentireorgan.Unfortunately,thesupplyofdonorheartscannotkeeppacewith
the demand; something must be done to enable us to repair damaged cardiac tissue and
generatewholeorganswhenneeded.Despitethefactthatdeathratesfromcoronaryheart
diseasearefalling,heartdiseaseremainstheleadingcauseofdeathworldwide.
Thisvolume,Cardiomyocytes:MethodsandProtocols,hasbeenassembledforscientists
interested in basic and applied biomedical research directed toward understanding the
development, genetics, and function of cardiomyocytes. The methods and protocols
contained herein address cell culture techniques, cardiomyocyte differentiation and redif-
ferentiation, experimental induction of cardiomyopathies, introducing genes into cardio-
myocytes,genomicapproachestotheunderstandingofcardiomyocytes,cryopreservation
ofneonatalcardiomyocytes,andmodelingofcardiomyocytefunction.Amongthechapters
of this work, readers will find complimentary areas of cardiomyocyte science that, taken
together,shouldinformindividualswithabroadrangeofinterests.
Thesecollectedcontributionswerewrittenbycurrentandnascentleadersinthefield
of cardiomyocyte biology. Together the authors have provided a wealth of methods that
canbeusedtofurtherexplorethemanyaspectsofcardiomyocytebiologythatweneedto
understand in order to better grasp the development and function of these cells and to
developthenextgenerationofeffectivetherapies.Thechaptersareorganizedthematically
with regard to cardiovascular disease, modelling of cardiomyocytes function, isolation of
cells,induceddifferentiationofcellsintocardiomyocytes,genetransferintocardiacmyo-
cytes,geneexpressionanalysis,andtheapplicationofnext-generationsequencingtoward
furtheringour understandingofcardiovasculardisease.
Of course it is impossible to include contributions from every researcher who is
contributing to this important field. Instead we have compiled a collection of chapters
thattogetherrepresentsomeoftheleadingandpotentiallymostimpactfulwork.Wehope
youfindtheminformativeandusefulinyourownlaboratories.
Rochester,NY,USA GaryR.Skuse
MaureenC.Ferran
v

Contents
Preface.................................................................... v
Contributors............................................................... ix
1 GeneratingPrimaryCulturesofMurineCardiacMyocytes
andCardiacFibroblaststoStudyViralMyocarditis ........................ 1
BarbaraSherry
2 EnrichmentofCardiomyocytesinPrimaryCulturesofMurine
NeonatalHearts....................................................... 17
SreejitParameswaran,RajalakshmiSanthakumar,
PrasannaVidyasekar,andRamaS.Verma
3 DeepSequencingofCardiacMicroRNA-mRNAInteractomes
inClinicalandExperimentalCardiomyopathy ............................ 27
ScotJ.MatkovichandGeraldW.DornII
4 Next-GenerationSequencingTechnologyintheGenetics
ofCardiovascularDisease............................................... 51
CeciliaVecoli
5 ComputationalCardiacElectrophysiology:ImplementingMathematical
ModelsofCardiomyocytestoSimulateActionPotentialsoftheHeart....... 65
MichaelM.BellandElizabethM.Cherry
6 MethodsofMyofibrillogenesisModeling................................. 75
NancyK.DrewandAnnaGrosberg
7 UsingtheMechanicalBidomainModeltoAnalyzetheBiomechanical
BehaviorofCardiomyocytes............................................ 93
BradleyJ.Roth
8 FabricationofaMyocardialPatchwithCellsDifferentiated
fromHuman-InducedPluripotentStemCells ............................ 103
LeiYe,JoydeepBasu,andJianyiZhang
9 EfficientDifferentiationofCardiomyocytesfromHumanPluripotent
StemCellswithGrowthFactors......................................... 115
RajneeshJha,Ren-HeXu,andChunhuiXu
10 Isolation,Culturing,andCharacterizationofCardiacMuscleCells
fromNonhumanPrimateHeartTissue .................................. 133
StevenM.HoynowskiandJohnW.Ludlow
11 MouseEmbryonicStemCell-DerivedCardiacMyocytes
inaCellCultureDish.................................................. 145
CarleyGlass,ReetuSingla,AnshuArora,andDinenderK.Singla
12 CryopreservationofNeonatalCardiomyocytes............................ 153
AdamC.Vandergriff,M.TaylorHensley,andKeCheng
13 EvaluationofSarcomericOrganizationinHumanPluripotent
StemCell-DerivedCardiomyocytes...................................... 161
ChrishanJ.A.RamachandraandWinstonShim
vii
viii Contents
14 ElectrotonicCoupledMetabolicPurificationofChickCardiomyocytes ...... 167
WinstonShim,HaiyangYu,K.P.MyuMaiJa,
MuhammadParasuram,KeePahLim,andPhilipWong
15 GeneTransferintoCardiacMyocytes.................................... 177
SarahE.LangandMargaretV.Westfall
16 Analysisof4DMyocardialWallMotionDuringEarlyStages
ofChickHeartDevelopment........................................... 191
MadelineMidgettandSandraRugonyi
Index..................................................................... 213
Contributors
ANSHUARORA (cid:1) BiomolecularScienceCenter,BurnettSchoolofBiomedicalSciences,
CollegeofMedicine,UniversityofCentralFlorida,Orlando,FL,USA
JOYDEEPBASU (cid:1) TissueEngineeringandAssayDevelopment,Zen-Bio,Inc.,Research
TrianglePark,NCUSA;ProcessResearchandTranslation,Tengion,Inc.,
Winston-Salem,NC,USA
MICHAELM.BELL (cid:1) SchoolofMathematicalSciences,RochesterInstituteofTechnology,
Rochester,NY,USA
KECHENG (cid:1) DepartmentofMolecularBiomedicalSciences,CollegeofVeterinaryMedicine,
NorthCarolinaStateUniversity,Raleigh,NC,USA;UNC/NCSUJointDepartmentof
BiomedicalEngineering,ChapelHill,NC,USA
ELIZABETHM.CHERRY (cid:1) SchoolofMathematicalSciences,RochesterInstituteofTechnology,
Rochester,NY,USA
GERALD W.DORN II (cid:1) Center forPharmacogenomics,DepartmentofInternalMedicine,
WashingtonUniversitySchoolofMedicine,St.Louis,MO,USA
NANCY K.DREW (cid:1) TheEdwardsLifesciencesCenter forAdvancedCardiovascular
Technology,Irvine,CA,USA
CARLEYGLASS (cid:1) BiomolecularScienceCenter,BurnettSchoolofBiomedicalSciences,
CollegeofMedicine,UniversityofCentralFlorida,Orlando,FL,USA
ANNAGROSBERG (cid:1) TheEdwardsLifesciencesCenter forAdvancedCardiovascular
Technology,Irvine,CA,USA
M.TAYLORHENSLEY (cid:1) DepartmentofMolecularBiomedicalSciences,CollegeofVeterinary
Medicine,NorthCarolinaStateUniversity,Raleigh,NC,USA
STEVENM.HOYNOWSKI (cid:1) ZenBio,Inc.,ResearchTrianglePark,NC,USA
K.P.MYUMAIJA (cid:1) StemCellLaboratory,NationalHeartResearchInstituteSingapore
(NHRIS),NationalHeartCentreSingapore(NHCS),Singapore,Singapore
RAJNEESHJHA (cid:1) DepartmentofPediatrics,EmoryUniversitySchoolofMedicineand
Children’sHealthcareofAtlanta,Atlanta,GA,USA
SARAH E.LANG (cid:1) PrograminCellularandMolecularBiology,UniversityofMichigan,
AnnArbor,MI,USA
KEEPAHLIM (cid:1) StemCellLaboratory,NationalHeartResearchInstituteSingapore
(NHRIS),NationalHeartCentreSingapore(NHCS),Singapore,Singapore
JOHNW.LUDLOW (cid:1) RegenerativeMedicine,ZenBio,Inc.,ResearchTrianglePark,NC,
USA
SCOTJ.MATKOVICH (cid:1) Center forPharmacogenomics,DepartmentofInternalMedicine,
WashingtonUniversitySchoolofMedicine,St.Louis,MO,USA
MADELINEMIDGETT (cid:1) DepartmentofBiomedicalEngineering,OregonHealth&Science
University,Portland,OR,USA
SREEJITPARAMESWARAN (cid:1) StemCellandMolecularBiologyLaboratory,
DepartmentofBiotechnology,IndianInstituteofTechnology,Chennai,
TamilNadu,India
MUHAMMADPARASURAM (cid:1) StemCellLaboratory,NationalHeartResearchInstitute
(NHRIS)Singapore,NationalHeartCentreSingapore (NHCS),Singapore,Singapore
ix