Human Embryonic Stem Cell Protocols

415 Pages·2015·11.988 MB·English

by  Kursad Turksen

Methods in
Molecular Biology 1307
Kursad Turksen Editor
Human
Embryonic Stem
Cell Protocols
Third Edition
M M B
ETHODS IN OLECULAR IOLOGY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire,AL109AB,UK
Forfurther volumes: http://www.springer.com/series/7651
.
Human Embryonic Stem
Cell Protocols
Third Edition
Edited by
Kursad Turksen
Ottawa Hospital Research Institute
Ottawa, ON, Canada
Editor
KursadTurksen
OttawaHospitalResearchInstitute
Ottawa,ON,Canada
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-4939-2667-1 ISBN978-1-4939-2668-8 (eBook)
DOI10.1007/978-1-4939-2668-8
LibraryofCongressControlNumber:2015947127
SpringerNewYorkHeidelbergDordrechtLondon
#SpringerScience+BusinessMediaNewYork2016
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Preface
Inspiteofethicalandpoliticalconcernsregardingtheuseofhumanembryonicstemcells,
theirpotentialtoadvancenotonlyregenerativemedicineapplicationsbutalsoour funda-
mentalunderstandingofstemcellbiologycontinuestodriveinterestinresearchwiththese
cells. In both cases, delineating the steps in and identifying the molecules responsible for
thecommitmentanddifferentiationofthesepluripotentcellstodifferentcellfatesremains
crucial. With this in mind, I felt that it would be timely to collect some of the most
interesting and useful protocols that have emerged during the last several years. I am
therefore extremely pleased and thankful to those groups who agreed to share their
valuable protocols with others interested in exploring stem cell biology questions in a
researchsetting.
Onceagain,IwouldliketoacknowledgeJohnWalker,EditorinChiefoftheseries,for
continuously fostering my efforts in this series. I am also grateful to Patrick Marton,
Executive Editor of the series, for encouraging me throughout the editorial process. In
addition, I thank David Casey for keeping me on track and pointing out all those details
thatmakecontributionstotheseriesconsistentanddependable.
Ottawa,ON,Canada KursadTurksen
v

Contents
Preface.................................................................... v
Contributors............................................................... xi
DerivationofHumanEmbryonicStemCellLinesfromVitrified
HumanBlastocysts......................................................... 1
CaraK.Bradley,JuliaSchaft,TammieK.Roy,BiljanaDumevska,
andTeijaT.Peura
®
CharacterizingPluripotentStemCellsUsingtheTaqMan hPSC
ScorecardTMPanel......................................................... 25
JeffreyFergus,ReneQuintanilla,andUmaLakshmipathy
GrowthofHumanPluripotentStemCellsUsingFunctionalHuman
ExtracellularMatrix........................................................ 39
AndresSanz-Garcia,MiodragStojkovic,andCarmenEscobedo-Lucea
EfficientExpansionofDissociatedHumanPluripotentStemCells
UsingaSyntheticSubstrate................................................. 61
EihachiroKawase
MicroarrayApproachtoIdentifytheSignalingNetworkResponsible
forSelf-RenewalofHumanEmbryonicStemCells ............................ 71
NoboruSatoandAliBrivanlou
MonitoringStemnessinLong-TermhESCCulturesbyReal-TimePCR.......... 89
AmparoGala´nandCarlosSimo´n
StudyofGapJunctionsinHumanEmbryonicStemCells ...................... 105
AlicePe´bayandRaymondC.B.Wong
ImmunofluorescenceMicroscopyandmRNAAnalysisofHuman
EmbryonicStemCells(hESCs)IncludingPrimaryCiliaAssociated
SignalingPathways......................................................... 123
MajLineaVestergaard,AashirAwan,CarolineBeckerWarzecha,
SørenTvorupChristensen,andClausYdingAndersen
AnalysisofIntracellularCalciumSignalinginHumanEmbryonic
StemCells ................................................................ 141
AdriennPe´ntek,KatalinPa´szty,andA´gotaApa´ti
GeneticManipulationofHumanEmbryonicStemCells........................ 149
RachelEiges
GeneticModificationinHumanPluripotentStemCells
byHomologousRecombinationandCRISPR/Cas9System.................... 173
HaipengXue,JianboWu,ShenglanLi,MahendraS.Rao,
andYingLiu
UseofMulticolorFlowCytometryforIsolationofSpecificCell
PopulationsDerivingfromDifferentiatedHumanEmbryonicStemCells ........ 191
IsabellaMengarelli,AndrewFryga,andTizianoBarberi
vii
viii Contents
DefinitiveEndoderm DifferentiationofHumanEmbryonicStem
CellsCombinedwithSelectiveEliminationofUndifferentiatedCells
byMethionineDeprivation ................................................. 205
TomonoriTsuyama,NobuakiShiraki,andShoenKume
DerivationofEndothelialCellsandPericytesfromHuman
PluripotentStemCells ..................................................... 213
SravantiKusumaandSharonGerecht
DifferentiationofHumanEmbryonicStemCellsonPeriodontal
LigamentFibroblasts....................................................... 223
Y.MuratElc¸in,Bu¨lendI˙nanc¸,andA.EserElc¸in
DerivationofEpithelialCellsfromHumanEmbryonicStemCells
asanInVitroModelofVocalMucosa........................................ 237
VlastaLungova,CiaraLeydon,andSusanThibeault
HumanEmbryonicandHepaticStemCellDifferentiationVisualized
inTwoandThreeDimensionsBasedonSerialSections ........................ 245
PeterS.Vestentoft,ChristianB.Brøchner,NielsLynnerup,
ClausYdingAndersen,andKjeldMøllga˚rd
DerivationofChondrogenicCellsfromHumanEmbryonic
StemCellsforCartilageTissueEngineering................................... 263
WeiSeongTohandTongCao
MicrogroovedSurfaceModulatesNeuronDifferentiation
inHumanEmbryonicStemCells............................................ 281
DavidLu,Chi-ShuoChen,Chao-SungLai,SushantSoni,
TaranzeLam,ClarenceLe,EricY.-T.Chen,ThienNguyen,
andWei-ChunChin
DirectedDifferentiationofHumanEmbryonicStemCells
intoNeuralProgenitors .................................................... 289
ErinBandaandLauraGrabel
DirectConversionofPluripotentHumanEmbryonicStem
CellsUnderDefinedCultureConditionsintoHumanNeuronal
orCardiomyocyteCellTherapyDerivatives................................... 299
XuejunH.Parsons
GenerationofEpithelialCellPopulationsfromHuman
PluripotentStemCellsUsingaSmall-MoleculeInhibitor
ofSrcFamilyKinases....................................................... 319
JoshuaA.Selekman,XiaojunLian,andSeanP.Palecek
Dual-SMADInhibition/WNTActivation-BasedMethodstoInduce
NeuralCrestandDerivativesfromHumanPluripotentStemCells............... 329
StuartM.Chambers,YvonneMica,GabsangLee,LorenzStuder,
andMarkJ.Tomishima
AcceleratedThree-DimensionalNeuroepitheliumFormation
fromHumanEmbryonicStemCellsandItsUseforQuantitative
DifferentiationtoHumanRetinalPigmentEpithelium......................... 345
YuZhu,SvenSchreiter,andEllyM.Tanaka
Contents ix
EfficientProductionofPhotoreceptorPrecursorCellsfromHuman
EmbryonicStemCells...................................................... 357
AnatYanai,ChristopherLaver,AaronW.Joe,andKevinGregory-Evans
GenerationofMegakaryocytesandPlateletsfromHuman
PluripotentStemCells ..................................................... 371
MarjoriePick
ASimpleProtocolfor theGenerationofCardiomyocytesfromHuman
PluripotentStemCells ..................................................... 379
GlenLesterSequiera,AshishMehta,andWinstonShim
Erratumto:MicrogroovedSurfaceModulatesNeuronDifferentiation
inHumanEmbryonicStemCells............................................ 385
DavidLu,Chi-ShuoChen,Chao-SungLai,SushantSoni,
TaranzeLam,ClarenceLe,EricY.-T.Chen,ThienNguyen,
andWei-ChunChin
Index..................................................................... 387