Germline Stem Cells

243 Pages·2016·6.432 MB·English

by  Michael Buszczak (ed.)

Methods in
Molecular Biology 1463
Michael Buszczak Editor
Germline
Stem Cells
Second 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

Germline Stem Cells
Second Edition
Edited by
Michael Buszczak
Department of Molecular Biology
University of Texas Southwestern Medical Center
Dallas, TX, USA
Editor
MichaelBuszczak
DepartmentofMolecularBiology
UniversityofTexasSouthwesternMedicalCenter
Dallas,TX,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-4939-4015-8 ISBN978-1-4939-4017-2 (eBook)
DOI10.1007/978-1-4939-4017-2
LibraryofCongressControlNumber:2016945156
©SpringerScience+BusinessMediaNewYork2008,2017
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Preface
Adult stem cells maintain tissue homeostasis by producing progeny that replace cells lost
throughthecourseofnormalcellularturnoverorbecauseofinjury.Germlinestemcellsare
uniqueamongst stem cellsbecause theyproduce daughtercellsthat develop into gametes,
which ultimately serve to give rise to the next generation. Thus, germline stem cells are
essentialfor thecontinuedpropagationofmanysexuallyreproducingorganisms.
Germcellsareimbuedwithseveraluniqueproperties.Theyaretheonlycellstoundergo
meiosis. Germ cellsacross species express many of thesame markersand experienceexten-
siveepigeneticreprogramming.Thesecellshavespecializedmechanismsthathelpmaintain
the integrity of the genome and prevent the integration of selfish DNA elements. Several
somatictumorsexpressgenesnormallyspecifictogermcellsandgermcellsthemselvescan
giverisetospecifictypesofcancer.Therefore,abetterunderstandingofgermcellswillhave
abroadimpactacrossmanyfieldsofbiology.
Germline stem cell biology has witnessed an explosion of new findings and techniques
since that last edition of Germline Stem Cells in the Methods in Molecular Biology series.
The optimization of live-cell imaging techniques, improved cell purification protocols and
theidentificationofgermcellsinanumberofgeneticallytractableorganismsnowallowsfor
the characterization of germ cells at a greater depth and higher resolution than previously
attainable.Thisnew editionofGermlineStemCells isintended toprovideresearcherswith
selected genetic, molecular, biochemical, and cell biological techniques used in germ cell
research.Whilethefocushereisonprimordialgermcellsandgermlinestemcells,manyof
the techniques and principles presented in the chapters of this issue may be applicable to
manydifferenttypesofadultstemcells.
IwouldliketothankProf.JohnM.WalkerandthestaffatSpringerfortheirinvitation,
assistance, and patience during the preparation of this book. I would like to thank Nevine
Shalabyforherhelpthroughoutthisentireprocess.Iwouldalsoliketoexpressmysincere
appreciationandgratitudetothevariouscontributorsforsharingtheirinsightsandexpertise
withthegermlinestemcellresearchcommunity.
Dallas,TX,USA MichaelBuszczak
v

Contents
Preface ..................................................................... v
Contributors................................................................. ix
1 AnalysisoftheC.elegansGermlineStemCellPool.......................... 1
SarahL.Crittenden,HannahS.Seidel,andJudithKimble
2 MethodsforStudyingtheGermlineoftheHumanParasite
Schistosomamansoni..................................................... 35
JulieN.R.CollinsandJamesJ.CollinsIII
3 EvaluationoftheAsymmetricDivisionofDrosophila
MaleGermlineStemCells ............................................... 49
MayuInabaandYukikoM.Yamashita
4 LiveImagingoftheDrosophilaTestisStemCellNiche ...................... 63
LeahJ.GreenspanandErikaL.Matunis
5 RNAIsolationfromEarlyDrosophilaLarvalOvaries ........................ 75
DanaGanczandLilachGilboa
6 Live-CellImagingoftheAdultDrosophilaOvaryUsing
ConfocalMicroscopy.................................................... 85
NevineA.ShalabyandMichaelBuszczak
7 MeasurementofmRNAPoly(A)TailLengthsinDrosophila
FemaleGermCellsandGerm-LineStemCells ............................. 93
AymericChartier,WillyJoly,andMartineSimonelig
8 IdentificationofGerm-LineStemCellsinZebrafish......................... 103
BruceW.Draper
9 PrimordialGermCellIsolationfromXenopuslaevisEmbryos ................ 115
AmandaM.Butler,TristanAguero,KarenM.Newman,
andMaryLouKing
10 Single-CellLineageAnalysisofOogenesisinMice.......................... 125
LeiLeiandAllanC.Spradling
11 VisualizationandLineageTracingofPax7+Spermatogonial
StemCellsintheMouse................................................. 139
GinaM.Aloisio,IleanaCuevas,YujiNakada,ChristopherG.Pen˜a,
andDiegoH.Castrillon
12 TransplantationasaQuantitativeAssaytoStudyMammalian
MaleGermlineStemCells ............................................... 155
AileenR.HelselandJonM.Oatley
13 MouseFetalGermCellIsolationandCultureTechniques ................... 173
CassyM.Spiller,GuillaumeBurnet,andJosephineBowles
vii
viii Contents
14 RattusnorvegicusSpermatogenesisColony-Forming Assays.................. 185
F.KentHamra
15 IdentificationofMousepiRNAPathwayComponents
UsingAnti-MIWI2Antibodies........................................... 205
TakamasaHirano,HidetoshiHasuwa,andHaruhikoSiomi
16 EfficientInductionandIsolationofHumanPrimordial
GermCell-LikeCellsfromCompetentHuman
PluripotentStemCells................................................... 217
NaokoIrieandM.AzimSurani
Index ...................................................................... 227
Contributors
TRISTANAGUERO (cid:1) DepartmentofCellBiology,UniversityofMiamiMillerSchoolof
Medicine,Miami,FL,USA
GINAM.ALOISIO (cid:1) DepartmentofPathologyandCecilH.andIdaGreenCenter for
ReproductiveBiologySciences,UniversityofTexasSouthwesternMedicalCenter,Dallas,
TX,USA
JOSEPHINEBOWLES (cid:1) SchoolofBiomedicalSciences,TheUniversityofQueensland,Brisbane,
QLD,Australia
GUILLAUMEBURNET (cid:1) SchoolofBiomedicalSciences,TheUniversityofQueensland,Brisbane,
QLD,Australia
MICHAELBUSZCZAK (cid:1) DepartmentofMolecularBiology,UniversityofTexasSouthwestern
MedicalCenter,Dallas,TX,USA
AMANDAM.BUTLER (cid:1) DepartmentofCellBiology,UniversityofMiamiMillerSchoolof
Medicine,Miami,FL,USA
DIEGOH.CASTRILLON (cid:1) DepartmentofPathologyandCecilH.andIdaGreenCenter for
ReproductiveBiologySciences,UniversityofTexasSouthwesternMedicalCenter,Dallas,
TX,USA
AYMERICCHARTIER (cid:1) mRNARegulationandDevelopment,InstitutdeGe´ne´tiqueHumaine,
CNRSUPR1142andUniversityofMontpellier,Montpellier,Cedex5,France
JULIEN.R.COLLINS (cid:1) DepartmentofPharmacology,UniversityofTexasSouthwestern
MedicalCenterr,Dallas,TX,USA
JAMESJ.COLLINSIII (cid:1) DepartmentofPharmacology,UniversityofTexasSouthwestern
MedicalCenter,Dallas,TX,USA
SARAH L.CRITTENDEN (cid:1) DepartmentofBiochemistry,HowardHughesMedicalInstitute,
UniversityofWisconsin-Madison,Madison,WI,USA
ILEANA CUEVAS (cid:1) DepartmentofPathologyandCecilH.andIdaGreenCenter for
ReproductiveBiologySciences,UniversityofTexasSouthwesternMedicalCenter,Dallas,
TX,USA
BRUCEW.DRAPER (cid:1) DepartmentofMolecularandCellularBiology,Universityof
California,Davis,Davis,CA,USA
DANAGANCZ (cid:1) DepartmentofBiologicalRegulation,WeizmannInstituteofScience,
Rehovot,Israel
LILACH GILBOA (cid:1) DepartmentofBiologicalRegulation,WeizmannInstituteofScience,
Rehovot,Israel
LEAHJ.GREENSPAN (cid:1) DepartmentofCellBiology,JohnsHopkinsUniversitySchoolof
Medicine,Baltimore,MD,USA
F.KENTHAMRA (cid:1) DepartmentofPharmacology,CecilH.&IdaGreenCenter for
ReproductiveBiologySciences,UniversityofTexasSouthwesternMedicalCenter,Dallas,
TX,USA
HIDETOSHIHASUWA (cid:1) DepartmentofMolecularBiology,KeioUniversitySchoolofMedicine,
Tokyo,Japan
AILEEN R.HELSEL (cid:1) Center forReproductiveBiology,SchoolofMolecularBiosciences,
CollegeofVeterinaryMedicine,WashingtonStateUniversity,Pullman,WA,USA
ix