Bone Research Protocols

662 Pages·2019·19.696 MB·English

by  Aymen I. Idris

Methods in
Molecular Biology 1914
Aymen I. Idris Editor
Bone Research
Protocols
Third Edition
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire,AL109AB,UK
Forfurther volumes:
http://www.springer.com/series/7651
Bone Research Protocols
Third Edition
Edited by
Aymen I. Idris
Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK
Editor
AymenI.Idris
DepartmentofOncologyandMetabolism
UniversityofSheffield
Sheffield,UK
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-4939-8996-6 ISBN978-1-4939-8997-3 (eBook)
https://doi.org/10.1007/978-1-4939-8997-3
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Preface
Researchinthebonefieldhasadvancedatarapidratewithmajorstrideshavebeenmadein
the field of musculoskeletal genetics, endocrinology, osteoimmunology, rheumatology,
molecular and cell biology, and pharmacology, to name a few. For 15 years, and through
two editions, Springer Protocols has presented laboratory methods to researchers in this
area in an exceptionally appealing and informative manner. Thus, I was delighted to be
presented with the opportunity to participate in the revision of this trusted source of
accurate,provenboneresearchmethods.
ThethirdeditionofBoneResearchProtocolsincludesaplethoraoflaboratoryprotocols
that are intended for basic and clinical bone researchers. In the tradition of the Methods in
Molecular Medicine series, the protocols are presented in a “recipe” style that provides a
step-by-step description of procedures that experienced researcher and those who are
entering the bone field can take to the laboratory and immediately apply in their own
research. As with previous editions, protocols include a description of relevant theoretical
aspects of methods, related diagrams, images, flowchart, timelines, and tables, as well as
additional “Notes” sections to suggest alternative procedures and to highlight limitations
andpitfallsandhowtheycanbeavoided.
The content ofthe latest edition isorganized into five partsthat cover a widerange of
established and novel bone research procedures. The first part that includes methods for
isolation, generation, and analysis ofosteoclasts, osteoblasts,osteocytes, and stem cellshas
been updated, and a new chapter on 2D and 3D co-cultures of bone and cancer cells has
been added. A new section that covers ex vivo models of bone organ and cell co-culture
system has been added. Part 3 contains a number of chapters that focus on biochemical
procedures for isolation, purification, quantification, and analysis of protein, mRNA, and
DNAinbonecellsandachapteronhowtostudysignalingpathwaysinbonecells.
Mychallengeineditingthethirdeditionhasbeentowidenthescopeofthethirdpartof
the book by including a number of new protocols while maintaining the cohesive and
inviting style that has distinguished the preceding two editions. I, therefore, extensively
revisedPart4ofthebooktocoverawiderangeofanimalmodelsofboneremodelingthat
include standard protocols such as in vivo models for estrogen deficiency and mechanical
loading,aswellasanumberofnewchapters.Twoadditionalchaptersonrodentmodelsof
rheumatoidarthritisandosteoarthritishavebeenadded,areflectionontherecentexplosion
inresearchininflammationandarthritis.
Cancer-associated bone disease is another area of research that has recently become an
importanttomanybonescientists.Withthisinmind,Iincludedalistofnewcancer-related
chaptersthatcoverprimarybonecancersandbonemetastasisassociatedwithvariouscancers
includingbreast,prostate,multiplemyeloma,andmelanoma.Bonepainisanotherexpand-
ingareaofboneresearch,andanewchapteronassessmentofcancer-inducedbonepainhas
been added. This section is followed by the final part of this edition that includes an
extensive list of chapters that provide detailed description of most of the most frequently
usedboneanalyticalandimagingtechniques.
I am proud that the content of the third edition was written by a cross-generational
groupofearlyandadvancedcareerresearcherswithestablishedtrackrecordsonthefieldof
bone research. This unique process offers special support to young investigators by
v
vi Preface
providing a platform to publish laboratory methods in their particular area of expertise. I
hopethatanyoneinterestedinboneresearch,betheybasicscientistorclinician,willfindthe
extensivelistofprotocolsincludedinthiseditionuseful.
Finally, I would like to acknowledge the series editor Professor John Walker, previous
editors Professors Stuart H. Ralston and Miep Helfrich and the many bone researchers
whoseresearchhasmadeitpossibletodeveloptheexperimentalproceduresandtechniques
described.
Sheffield,UK AymenI.Idris
Contents
Preface ..................................................................... v
Contributors................................................................. xi
PART I IN VITRO BONE CELL CULTURES
1 IsolationandGenerationofOsteoclasts.................................... 3
NadiaRucci,AlbertaZallone,andAnnaTeti
2 IsolationandGenerationofOsteoblasts ................................... 21
IneˆsP.Perpe´tuo,LucieE.Bourne,andIsabelR.Orriss
3 Isolation,Purification,Generation,andCultureofOsteocytes................ 39
JonathanH.Gooi,LingYeongChia,ChristinaVrahnas,
andNatalieA.Sims
4 Isolation,Differentiation,andCharacterizationofHumanBone
MarrowStemCellsInVitroandInVivo................................... 53
JanosKanczler,RahulS.Tare,PatrickStumpf,TimothyJ.Noble,
CameronBlack,andRichardO.C.Oreffo
5 2Dand3DInVitroCo-CultureforCancerandBoneCell
InteractionStudies...................................................... 71
SilviaMarino,RyanT.Bishop,Dani¨elledeRidder,JesusDelgado-Calle,
andMichaelaR.Reagan
6 UsingCellandOrganCultureModelstoAnalyzeResponses
ofBoneCellstoMechanicalStimulation................................... 99
SoraiaP.Caetano-Silva,AstridNovicky,BehzadJavaheri,
SimonC.F.Rawlinson,andAndrewA.Pitsillides
PART II BIOCHEMICAL AND MOLECULAR ANALYSIS OF BONE CELLS
7 AnalysisofSignalingPathwaysbyWestern Blotting
andImmunoprecipitation................................................ 131
SilviaMarinoandAymenI.Idris
8 AnalysisofTranscriptionalRegulationinBoneCells ........................ 145
HuilinJin,AntoniaSophocleous,AsimAzfer,andStuartH.Ralston
9 AnalysisofmRNA,miRNA,andDNAinBoneCellsbyRT-qPCR
andInSituHybridization................................................ 169
BriceMoukengue,Je´roˆmeAmiaud,CamilleJacques,Ce´lineCharrier,
BenjaminOry,andFrancoisLamoureux
PART III EX VIVO MODELS
10 TheExVivoOrganCultureofBone...................................... 199
KatherineA.Staines,GenevieveBrown,andColinFarquharson
vii
viii Contents
11 Ex-VivoModelSystemsofCancer-BoneCellInteractions ................... 217
ErdjanSalih
12 MechanicalPropertiesofBoneExVivo.................................... 241
SimonR.GoodyearandRichardM.Aspden
PART IV IN VIVO BONE MODELS
13 Ovariectomy/OrchiectomyinRodents.................................... 261
AntoniaSophocleousandAymenI.Idris
14 InVivoModelsofRheumatoidArthritis................................... 269
BettinaGr¨otsch,AlineBozec,andGeorgSchett
15 OsteoarthritisMouseModelofDestabilizationoftheMedialMeniscus ....... 281
AntoniaSophocleousandCarmenHuesa
16 ModelsofProstateCancerBoneMetastasis................................ 295
SunHeePark,MatthewRobertEber,andYusukeShiozawa
17 AnimalModelsofBreastCancerBoneMetastasis........................... 309
ClaudiaTulotta,ArwinGroenewoud,B.EwaSnaar-Jagalska,
andPenelopeOttewell
18 MurineModelsofBoneSarcomas ........................................ 331
CamilleJacques,NathalieRenema,BenjaminOry,CarlR.Walkley,
AgamemnonE.Grigoriadis,andDominiqueHeymann
19 MouseModelsofMelanomaBoneMetastasis.............................. 343
O¨zgeUluc¸kan
20 AnimalModelsofMultipleMyeloma...................................... 349
SamW.Z.OlechnowiczandClaireM.Edwards
21 TheCalvarialInjectionAssay............................................. 361
RobertJ.van’tHof
22 InVivoModelsofMechanicalLoading.................................... 369
BehzadJavaheri,NathalieBravenboer,AstridD.Bakker,
AlbertvanderVeen,RobertoLopesdeSouza,LeanneSaxon,
andAndrewA.Pitsillides
23 AnimalModelsfor theStudyofBone-DerivedPain......................... 391
AustenL.Thompson,TallyM.Largent-Milnes,andToddW.Vanderah
PART V MICROSCOPICAL AND IMAGING TECHNIQUES
24 HistomorphometryinRodents........................................... 411
ReinholdG.ErbenandMartinGl¨osmann
25 ImmunostainingofSkeletalTissues ....................................... 437
AnkeJ.RoelofsandCosimoDeBari
26 Techniquesfor theStudyofApoptosisinBone............................. 451
ValMannandBrendonNoble
27 LiveCellImagingofBoneCellandOrganCultures ........................ 467
SarahL.Dallas,PatriciaA.Veno,andLeAnnM.Tiede-Lewis
Contents ix
28 AnalysisofBoneArchitectureinRodentsUsingMicro-Computed
Tomography ........................................................... 507
RobertJ.van’tHofandEnricoDall’Ara
29 BoneMeasurementsbyPeripheralQuantitativeComputed
TomographyinRodents................................................. 533
Ju¨rgAndreasGasserandJohannesWillnecker
30 QuantitativeX-RayImagingofMouseBonebyFaxitron .................... 559
NatalieC.Butterfield,JohnG.Logan,JulianWaung,
GrahamR.Williams,andJ.H.DuncanBassett
31 ScanningElectronMicroscopyofBone.................................... 571
AlanBoyde
32 TransmissionElectronMicroscopyofBone ................................ 617
VincentEverts,AnnekeNiehof,WikkyTigchelaar-Gutter,
andWouterBeertsen
33 TrackingCancerCellsColonizationinRodentBoneUsing
Two-PhotonMicroscopy ................................................ 631
GloriaAlloccaandNingWang
34 FourierTransformInfraredImagingofBone............................... 641
EleftheriosP.Paschalis
35 RamanMicroscopyandBone ............................................ 651
SimonR.GoodyearandRichardM.Aspden
Index ...................................................................... 661
Contributors
GLORIA ALLOCCA (cid:1) DepartmentofOncologyandMetabolism,TheMellanbyCentreforBone
Research,TheUniversityofSheffield,Sheffield,UK
JE´ROˆMEAMIAUD (cid:1) INSERM,UMR1238,BoneSarcomaandRemodelingofCalcifiedTissues,
Universite´deNantes,NantesAtlantiqueUniversite´s,Nantes,France
RICHARDM.ASPDEN (cid:1) ArthritisandMusculoskeletalMedicine,InstituteofMedicalSciences,
UniversityofAberdeen,Aberdeen,UK
ASIMAZFER (cid:1) RheumaticDiseasesUnit,CentreforGenomicsandExperimentalMedicine,
MRCIGMM,WesternGeneralHospital,UniversityofEdinburgh,Edinburgh,UK
ASTRIDD.BAKKER (cid:1) DepartmentofOralCellBiology,AcademicCentreforDentistry
Amsterdam,Amsterdam,TheNetherlands
J.H.DUNCANBASSETT (cid:1) MolecularEndocrinologyLaboratory,DepartmentofMedicine,
ImperialCollegeLondon,HammersmithHospitalCampus,London,UK
WOUTER BEERTSEN (cid:1) DepartmentofOralCellBiology,AcademicCentreforDentistry
Amsterdam(ACTA),UniversityofAmsterdam,VUUniversityAmsterdam,Amsterdam,
TheNetherlands;ResearchInstituteMOVE,VUUniversityAmsterdam,Amsterdam,The
Netherlands;DepartmentofPeriodontology,AcademicCentreforDentistryAmsterdam
(ACTA),UniversityofAmsterdam,Amsterdam,TheNetherlands
RYANT.BISHOP (cid:1) DepartmentofOncologyandMetabolism,MedicalSchool,Universityof
Sheffield,Sheffield,UK
CAMERON BLACK (cid:1) BoneandJointResearchGroup,CentreforHumanDevelopment,Stem
CellsandRegeneration,HumanDevelopmentandHealth,InstituteofDevelopmental
Sciences,UniversityofSouthampton,FacultyofMedicine,Southampton,UK
LUCIE E.BOURNE (cid:1) DepartmentofComparativeBiomedicalSciences,RoyalVeterinary
College,London,UK
ALANBOYDE (cid:1) DentalPhysicalSciences,BiophysicsSection,OralGrowthandDevelopment,
DentalInstitute,BartsandTheLondonSchoolofMedicineandDentistry,QueenMary
UniversityofLondon,London,UK
ALINE BOZEC (cid:1) DepartmentofMedicine3,RheumatologyandImmunology,University
HospitalErlangen,Erlangen,Germany
NATHALIEBRAVENBOER (cid:1) DepartmentClinicalChemistry,VUUniversityMedicalCenter,
Amsterdam,TheNetherlands;Center forBoneQualityandDepartmentofInternal
Medicine,DivisionEndocrinology,LeidenUniversityMedicalCenter,Leiden,The
Netherlands
GENEVIEVEBROWN (cid:1) DepartmentofBiomedicalEngineering,ColumbiaUniversity,
NewYork,NY,USA
NATALIEC.BUTTERFIELD (cid:1) MolecularEndocrinologyLaboratory,DepartmentofMedicine,
ImperialCollegeLondon,HammersmithHospitalCampus,London,UK
SORAIAP.CAETANO-SILVA (cid:1) SkeletalBiologyGroup,ComparativeBiomedicalSciences,The
RoyalVeterinaryCollege,London,UK
CE´LINE CHARRIER (cid:1) INSERM,UMR1238,BoneSarcomaandRemodelingofCalcified
Tissues,Universite´deNantes,NantesAtlantiqueUniversite´s,Nantes,France
xi