Hematopoietic Stem Cells: Methods and Protocols

284 Pages·2022·9.741 MB·English

by  Louis M. Pelus, Jonathan Hoggatt

Methods in
Molecular Biology 2567
Louis M. Pelus
Jonathan Hoggatt Editors
Hematopoietic
Stem Cells
Methods and Protocols
M M B
ETHODS IN OLECULAR IO LO GY
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JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
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Hematopoietic Stem Cells
Methods and Protocols
Edited by
Louis M. Pelus
Department of Microbiology & Immunology and Department of Medicine/Hematology
Oncology, Indiana University School of Medicine, Indianapolis, IN, USA
Jonathan Hoggatt
Moderna Therapeutics, Cambridge, MA, USA
Editors
LouisM.Pelus JonathanHoggatt
DepartmentofMicrobiology& ModernaTherapeutics
ImmunologyandDepartment Cambridge,MA,USA
ofMedicine/Hematology
Oncology,IndianaUniversity
SchoolofMedicine
Indianapolis,IN,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-0716-2678-8 ISBN978-1-0716-2679-5 (eBook)
https://doi.org/10.1007/978-1-0716-2679-5
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Cover Illustration Caption: The cover photo shows trilineage differentiation of expanded human bone marrow
MesenchymalStem/StromalCells(MSC)toOsteoblasts(left),Adipocytes(Middle)andChondrocytes(right). MSC
areintegralcomponentsofthehematopoieticnichethatnurtureandeducateHematopoieticStemCells.Osteogenic
differentiationwasdeterminedbystainingformineralizationofextracellularmatrixandcalciumdepositsusingAlizarin
Red S. Adipocytes were identified by production of lipid droplets using Oil Red O staining. Alcian blue staining of
glycosaminoglycansdeterminedchondrocytedifferentiation.ThecoverphotoisprovidedbyDr.PratibhaSingh(Indiana
UniversitySchoolofMedicine,Indianapolis,IN,USA).
ThisHumanaimprintispublishedbytheregisteredcompanySpringerScience+BusinessMedia,LLC,partofSpringer
Nature.
Theregisteredcompanyaddressis:1NewYorkPlaza,NewYork,NY10004,U.S.A.
Preface
Thefieldofexperimentalhematology,orhematopoiesisorhematopoieticstemcellbiology
aswemostlyrefertoittoday,hasgrowntremendouslyandcontinuestogrowspurredonby
ever-increasing technology and sophistication. Having entered the field in the early 1970s
(LMPelus),Iwasamazedbytheabilityoftechniquestovisualizethegrowthofindividual
bone marrow cells into colonies of myeloid and erythroid cells when given magical organ-
conditioned medium and the utility to evaluate responses to exogenously added entities.
Lethally irradiated mice could be salvaged by reintroduction of cells that formed spleen
nodules.Thepoorman’ssortingexperimentswherebylineagespecificitycouldbeobserved
and altered by separating cloned daughter cells, and the long-term culture systems that
allowedforevaluationofhematopoieticdifferentiationprovidedsomebasicunderstanding
ofstemandprogenitorcellfunction.
We have come so far from then. The hematopoietic hormones were cloned and made
readily available. FACS machines were developed for analysis and selection of single cells.
Monoclonalantibodiesanduniquefluorochromesandmasscytometryweredevelopedthat
allow us to use these machines to quantify stem and progenitor cell populations and
signaling pathways and transcription factors in these cells. Genetic tools to turn on, turn
off, or delete genes in living organisms were developed, as were single cell genomics and
proteomics.Insitumethodsletuspeerintothelivingbonemarrow.Newgeneticorganisms
and more relevant animal models continue to emerge to evaluate true in vivo function of
stemandprogenitorcellsandvalidatethefindingsuncoveredwithallthesetools.Withthese
tools,wehaveagreaterunderstandingofhematopoiesisandcellfunction,andnewtechno-
logicaladvancesoffer thepromisetolearnmore.
Previouseditionsofthisseriescoveringhematopoieticstemcellprotocolshaveprovided
thorough coverage of many developments in the field, focused on user friendlyand detail-
oriented format, to allow those inexperienced or new to the areas apply these methods to
their research and take advantage of the exceptional guidance offered by expert faculty. In
this new edition, our aims were to update technical advances in areas covered in previous
editions and incorporate new techniques focused on the molecular/genetic, cellular, and
whole organism levels. We were also keen to incorporate methods that apply stress to
hematopoiesis,whichwebelieveisamajorpathwaytounderstandhematopoieticstemcell
biology and drive research in many unexpected and exciting ways in the future. While not
exhaustivebyanymeans,thiseditionnowcontainschaptersfocusedonbetter understand-
ingtheroleofhematopoieticnichesandtheircellularcomponents.Lastly,wehaveincluded
chaptersdedicatedtoinvivomodelsthattestandquantitatestemcellfunctionandarekey
tofurtherdevelopmentoftherapeuticapplications.
Thisbookwouldnotbepossiblewithoutthetimeandeffortofallthecontributingauthors.
Theirdedicationtotheintentofthisseriesisoutstanding,andwethankthemfortheirsupport.
Wehopethatthisresourcewillbeavaluedadditiontoalllaboratoriesfocusedonunderstanding
hematopoieticstemcellbiologyandthetherapeuticadvancesthatcanbederived.
Indianapolis,IN,USA LouisM.Pelus
Cambridge,MA,USA JonathanHoggatt
v
Contents
Preface ..................................................................... v
Contributors................................................................. ix
PART I OVERVIEW
1 TheFactoryofBloodProduction:HematopoieticStemCells ........ ........ 3
JonathanHoggattandLouisM.Pelus
PART II MOLECULAR MODIFICATION AND EVALUATION
2 DetectionofDNADamageinHematopoieticStemCells..... ....... ........ 11
SaipriyaAyyarandIsabelBeerman
3 EvaluatingHistoneAcetylationinMouseHematopoieticStem
andProgenitorCellsUsingChromatinImmunoprecipitation ........ ........ 29
LiqiongLiu
4 CRISPRGeneEditingofHematopoieticStemandProgenitorCells.......... 39
RezaShahbazi,PatriciaLipson,KarthikeyaS.V.Gottimukkala,
DanielD.Lane,andJenniferE.Adair
5 LentiviralTransductionofNonhumanPrimateHematopoieticStem
andProgenitorCells ....... ........ ....... ....... ........ ....... ...... .. 63
ChuanfengWu,SoGunHong,AylinBonifacino,
andCynthiaE.Dunbar
PART III CELLULAR ISOLATION AND CHARACTERIZATION
6 IdentificationofNonhumanPrimateHematopoieticStemandProgenitor
Cells....... ........ ....... ........ ....... ....... ........ ....... ........ 87
StefanRadtkeandHans-PeterKiem
7 IsolationandCharacterizationofFetalLiverHematopoieticStemCells ....... 99
DiegoA.L(cid:1)opezandAnnaE.Beaudin
8 UtilizingCyTOFtoExamineHematopoieticStemandProgenitor
Phenotype ......... ....... ........ ....... ... .... .... .... ....... ........ 113
SafaF.MohamadandMaeganL.Capitano
9 HematopoieticStemCellIdentificationPostirradiation....... ....... ........ 127
AndreaM.Patterson,ChristieM.Orschell,andLouisM.Pelus
PART IV THE HEMATOPOIETIC NICHE AND ACCESSORY CELLS
10 IntravitalMicroscopyforHematopoieticStudies..... ........ ....... ........ 143
MyriamL.R.HaltalliandCristinaLoCelso
vii
viii Contents
11 LaserMicromachiningofBoneasaToolforStudyingBone
MarrowBiology .... ....... ........ ....... ....... ........ ....... ........ 163
ChristaHaase,DmitryRichter,andCharlesP.Lin
12 MSCandHSPCCoculture:MimickingExVivoBoneMarrowNiche......... 181
PratibhaSingh
13 IsolationofThymusStromalCellsfromHumanandMurineTissue .......... 191
KarinGustafssonandDavidT.Scadden
PART V TRANSPLANTATION AND IN VIVO MODELS
14 ExperimentalModelsofMouseandHumanHematopoieticStemCell
Transplantation..... ....... ........ ....... ....... .... .... ..... .. ........ 205
ScottH.Cooper,MaeganL.Capitano,andHalE.Broxmeyer
15 HematopoieticStemandProgenitorCellIdentification
andTransplantationinZebrafish..... ....... ....... ........ ....... ........ 233
EllenFraint,PengLv,FengLiu,TeresaV.Bowman,
andOwenJ.Tamplin
16 EstablishingaMurineModeloftheHematopoieticAcuteRadiation
Syndrome.......... ....... ........ ....... ....... ........ ....... ....... . 251
P.ArturPlett,LouisM.Pelus,andChristieM.Orschell
17 PurinergicSignalingandItsRoleinMobilizationofBoneMarrow
StemCells ......... ....... ........ ....... ....... ........ ....... ........ 263
MalwinaSuszynska,MateuszAdamiak,ArjunThapa,
MonikaCymer,JaninaRatajczak,MagdalenaKucia,
andMariuszZ.Ratajczak
Index ...................................................................... 281
Contributors
JENNIFERE.ADAIR • FredHutchinsonCancerCenter,Seattle,WA,USA;Universityof
Washington,Seattle,WA,USA
MATEUSZADAMIAK • DepartmentofRegenerativeMedicine,WarsawMedicalUniversity,
Warsaw,Poland
SAIPRIYAAYYAR • EpigeneticsandStemCellUnit,TranslationalGerontologyBranch,
NationalInstituteonAging,Baltimore,MD,USA
ANNAE.BEAUDIN • DepartmentofInternalMedicine,DivisionofHematologyand
HematologicMalignancies,UniversityofUtahSchoolofMedicine,SaltLakeCity,UT,
USA
ISABEL BEERMAN • EpigeneticsandStemCellUnit,TranslationalGerontologyBranch,
NationalInstituteonAging,Baltimore,MD,USA
AYLINBONIFACINO • TranslationalStemCellBiologyBranch,NationalHeart,Lung,and
BloodInstitute,NationalInstitutesofHealth,Bethesda,MD,USA
TERESAV.BOWMAN • GottesmanInstituteforStemCellBiologyandRegenerativeMedicine,
DepartmentofDevelopmentalandMolecularBiology,andDepartmentofMedicine
(Oncology),AlbertEinsteinCollegeofMedicineandMontefioreMedicalCenter,Bronx,
NY,USA
HALE.BROXMEYER • DepartmentofMicrobiology&Immunology,IndianaUniversity
SchoolofMedicine,Indianapolis,IN,USA
MAEGANL.CAPITANO • DepartmentofMicrobiologyandImmunology,IndianaUniversity
SchoolofMedicine,Indianapolis,IN,USA
CRISTINALOCELSO • ImperialCollegeLondon,London,UK;TheFrancisCrickInstitute,
London,UK
SCOTTH.COOPER • DepartmentofMicrobiology&Immunology,IndianaUniversitySchool
ofMedicine,Indianapolis,IN,USA
MONIKACYMER • DepartmentofRegenerativeMedicine,WarsawMedicalUniversity,
Warsaw,Poland
CYNTHIAE.DUNBAR • TranslationalStemCellBiologyBranch,NationalHeart,Lung,and
BloodInstitute,NationalInstitutesofHealth,Bethesda,MD,USA
ELLENFRAINT • DepartmentofPediatrics(PediatricHematology/OncologyandCellular
Therapy)andDepartmentofDevelopmentalandMolecularBiology,AlbertEinstein
CollegeofMedicineandMontefioreMedicalCenter,Bronx,NY,USA
KARTHIKEYAS.V.GOTTIMUKKALA • FredHutchinsonCancerCenter,Seattle,WA,USA
KARINGUSTAFSSON • Center forRegenerativeMedicine,MassachusettsGeneralHospital,
Boston,MA,USA;HarvardStemCellInstitute,Cambridge,MA,USA;Departmentof
StemCellandRegenerativeBiology,HarvardUniversity,Cambridge,MA,USA
CHRISTAHAASE • WellmanCenter forPhotomedicine,MassachusettsGeneralHospitaland
HarvardMedicalSchool,Boston,MA,USA;Center forSystemsBiology,Massachusetts
GeneralHospitalandHarvardMedicalSchool,Boston,MA,USA
MYRIAML.R.HALTALLI • ImperialCollegeLondon,London,UK;TheFrancisCrick
Institute,London,UK;Wellcome–MedicalResearchCouncilCambridgeStemCell
Institute,UniversityofCambridge,Cambridge,UK
JONATHANHOGGATT • ModernaTherapeutics,Cambridge,MA,USA
ix
x Contributors
SOGUNHONG • TranslationalStemCellBiologyBranch,NationalHeart,Lung,andBlood
Institute,NationalInstitutesofHealth,Bethesda,MD,USA
HANS-PETERKIEM • StemCellandGeneTherapyProgram,FredHutchinsonCancer
ResearchCenter,Seattle,WA,USA;DepartmentofMedicine,UniversityofWashington
SchoolofMedicine,Seattle,WA,USA;DepartmentofPathology,UniversityofWashington
SchoolofMedicine,Seattle,WA,USA
MAGDALENAKUCIA • StemCellInstitute,JamesGrahamBrownCancerCenter,Universityof
Louisville,Louisville,KY,USA;DepartmentofRegenerativeMedicine,WarsawMedical
University,Warsaw,Poland
DANIELD.LANE • FredHutchinsonCancerCenter,Seattle,WA,USA
CHARLESP.LIN • WellmanCenter forPhotomedicine,MassachusettsGeneralHospitaland
HarvardMedicalSchool,Boston,MA,USA;Center forSystemsBiology,Massachusetts
GeneralHospitalandHarvardMedicalSchool,Boston,MA,USA;HarvardStemCell
Institute,Cambridge,MA,USA
PATRICIALIPSON • FredHutchinsonCancerCenter,Seattle,WA,USA
FENGLIU • StateKeyLaboratoryofMembraneBiology,InstituteofZoology,Chinese
AcademyofSciences,Beijing,China;InstituteforStemCellandRegeneration,Chinese
AcademyofSciences,Beijing,China;UniversityofChineseAcademyofScience,Beijing,
China
LIQIONGLIU • DepartmentofMicrobiologyandImmunology,IndianaUniversitySchoolof
Medicine,Indianapolis,IN,USA;TriArmTherapeutics,Shanghai,PRChina
DIEGOA.LO´PEZ • DepartmentofPathology,UniversityofUtahSchoolofMedicine,SaltLake
City,UT,USA
PENGLV • StateKeyLaboratoryofMembraneBiology,InstituteofZoology,ChineseAcademy
ofSciences,Beijing,China;InstituteforStemCellandRegeneration,ChineseAcademyof
Sciences,Beijing,China;UniversityofChineseAcademyofScience,Beijing,China
SAFAF.MOHAMAD • DepartmentofHematologyandOncology,BostonChildren’sHospital/
HarvardSchoolofMedicine,Boston,MA,USA
CHRISTIEM.ORSCHELL • DepartmentofMedicine/HematologyOncology,Indiana
UniversitySchoolofMedicine,Indianapolis,IN,USA
ANDREA M.PATTERSON • DepartmentofMedicine/HematologyOncology,Indiana
UniversitySchoolofMedicine,Indianapolis,IN,USA
LOUISM.PELUS • DepartmentofMicrobiology&ImmunologyandDepartmentof
Medicine/HematologyOncology,IndianaUniversitySchoolofMedicine,Indianapolis,IN,
USA
P.ARTURPLETT • DepartmentofMedicine/HematologyOncology,IndianaUniversitySchool
ofMedicine,Indianapolis,IN,USA
STEFANRADTKE • StemCellandGeneTherapyProgram,FredHutchinsonCancerResearch
Center,Seattle,WA,USA
JANINARATAJCZAK • StemCellInstitute,JamesGrahamBrownCancerCenter,Universityof
Louisville,Louisville,KY,USA
MARIUSZZ.RATAJCZAK • StemCellInstitute,JamesGrahamBrownCancerCenter,
UniversityofLouisville,Louisville,KY,USA;DepartmentofRegenerativeMedicine,
WarsawMedicalUniversity,Warsaw,Poland