温医大生殖医学课件06干细胞

BasicKnowledgesaboutStemCells(干细胞基本知识) A.Definition

B.Classification

C.Characterization2.StemCellsinTestis(睾丸干细胞) A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary(卵巢干细胞）

A.Germlinestemcells

B.Somaticstemcells4.FutureDirections(未来方向) A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本课内容Stemcellsarethecellsthatcandivideessentiallywithoutlimittoformothercells.(无限分裂并形成其他细胞的细胞）Self-renewaltomaintainthemselves

(自我增殖)2)Differentiationtoformspecializedcells

(分化成子代细胞）Whatarestemcells?全能干细胞

(Totipotent

stemcells):Cells

canformallthecelltypesinabody,plustheextraembryonic,orplacental,cells.Embryoniccellswithinthefirstcoupleofcelldivisionsafterfertilizationaretheonlycellsthataretotipotent

(包括胎盘).多能干细胞

(Pluripotent

stemcells):Cellscangiverisetoallofthecelltypesthatmakeupthebody;embryonicstemcellsareconsideredpluripotent

(不包括胎盘).亚多能干细胞

(Multipotent

stemcells):Cellscandevelopintomorethanonecelltype,butaremorelimitedthanpluripotentcells;adultstemcellsandcordbloodstemcellsareconsideredmultipotent.单能干细胞

(Unipotent

stemcells):Cellreferstoacellthatcandifferentiatealongonlyonelineage.干细胞的多能性Totipotent滋养层囊胚Multipotentstemcells（亚多能干细胞）骨髓祖细胞淋巴祖细胞

1.Embryonicstemcells(胚胎干细胞）

Generatedfromcellsofapreimplantation-stageembryo

Totipotent

Pluripotent

2.“Somatic”or“adult”stemcells(成体干细胞)

Generatedfromdevelopingorwell-developedoradultorgans

Multipotent

Unipotent3.Inducedpluripotentstemcells(iPSCs)

(诱导多潜能干细胞) Generatedfromtransductionof4transcriptionalfactors(Oct4 (Pou5f1),Sox2,cMyc,andKlf4)

PluripotentClassification(分类）干细胞小巢及干细胞对称性和非对称性分裂干细胞小巢Stem-cellnicheThespecificanatomiclocationwherestemcellslive,whichcontainsextracelluarmatrixandregulatorycells.Theniche(home)playsimportantrolesindeterminingstemcells’fate.Theword'niche'oftenbereferedtoasstem-cellmicroenvironmentinvivoorinvitro.小巢是指干细胞“生活”的位置，其中包含细胞外基质和调控细胞。小巢（家）在调控干细胞的活动及确定干细胞的命运中起着重要的作用。“小巢”一词通常被泛指体内或体外干细胞生存的微环境。“归巢”是指干细胞从身体的其它位置，向其小巢回归的过程。BasicKnowledgesaboutStemCells A.Definition

B.Classification

C.CharacterizationStemCellsinTestis A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary

A.Germlinestemcells B.Somaticstemcells4.FutureDirections A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本课内容SPERMATOGENESIS:/file/view/spermatogenesis.jpg/298006750/800x446/spermatogenesis.jpg曲细精管间质GermcellsSertolicellsLeydigcellsMyoidcellsMacrophagesBloodvesselsSPERMATOGENESIS:/file/view/spermatogenesis.jpg/298006750/800x446/spermatogenesis.jpg/file/view/spermatogenesis.jpg/298006750/800x446/spermatogenesis.jpgDifferencesBetweenRodentsandHumanMOUSEHUMANGfra1c-KitID4PAX7BMI1TheAsmodelforspermatogonialmultiplicationandstemcellrenewal.DirkG.deRooijDevelopment2017;144:3022-3030©2017.PublishedbyTheCompanyofBiologistsLtd简单模型Thefragmentationmodelforspermatogonialmultiplicationandstemcellrenewal.DirkG.deRooijDevelopment2017;144:3022-3030©2017.PublishedbyTheCompanyofBiologistsLtd碎片模型ThehierarchicalAsmodel.DirkG.deRooijDevelopment2017;144:3022-3030©2017.PublishedbyTheCompanyofBiologistsLtd分级模型Gfra1C-KitSpermatogoniastemcellmarkersTruestemmarkersID4:inhibitorofdifferentiation4PAX7:pairedbox7BMI1:Bcell-specificMoloneymurine leukaemiavirusintegrationsite1DifferentatingmarkersNgn3:neurogenin3Gfra1:GDNFfamilyreceptoralpha1Fbxw7:Skp1-Cullin-F-box(SCF)-type ubiquitinligase

SummarySpermatogoniastemcells1.Highturnovercells,activeduringtheindividual’slifetime2.Numberoftheprogeniescanbegeneratedbyeachstemcellarelimited3.Rodentsgenerate500foldsofgermcellsthanhumanforeachstem

celldivision4.Threemodelshavebeenproposedbywhichhowspermatogoniastemcellsundergodifferentiation:Asmodel(简单模型);Fragmentationmodel(碎片模型);Hierarchicalmodel(分级模型)。TesticularStemCellsforSomaticcells

Sertolicells?(notyet)

Myiodcells?(notyet)

Leydigcells?Yes(PeritubularandPerivescularlocations)CD90绿色--管周细胞(smactin)红色—间质细胞(P450scc)

培养4周后，在管周myoid细胞的外表面，出现间质细胞BasicKnowledgesaboutStemCells A.Definition

B.Classification

C.CharacterizationStemCellsinTestis A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary

A.Germlinestemcells B.Somaticstemcells4.FutureDirections A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本课内容植物根尖生长(干细胞功能及顶尖生长）低等动物的卵巢含有生殖干细胞Celegans线虫（含生殖干细胞）Drosophila果蝇（含生殖干细胞）Medaka日本淡水鱼（含生殖干细胞）哺乳动物卵巢含有生殖干细胞吗？传统的观点：卵巢不含生殖干细胞Eggnumberislimitedandcouldbedepletedbyageof50JonathanL.Tilly2004哺乳动物成年卵巢含有生殖干细胞(Germline

Stem

Cells:

GSC)

JonathanL.TillyGroupNature2004;428:145–50.Countingatreticfollicles（33%

immature

follicles

are

atretic,whichcouldresultindepletionoffolliclesmuchearlier)

（卵泡萎缩速度太快）GermcellmarkersVasahomologue(MVH,aks

Deadbox4,orDDX4)andsynaptonemalcomplexprotein3(SCP3)-positivecellswerefoundatovariansurfaceepithelium(previouslytermedgerminal

epithelium).

BrdU

incorporation

of

MVH-positive

cells（卵泡外表达生殖细胞基因的细胞）Germ

cell

toxicant

(busulfan)onlyaffectedfolliclein2weeks,suggestingitaffectedGSC（毒物影响卵泡的速度太慢）OccurrenceofGFP-expressingoocytessurroundedbynon-GFPovariescellsaftertransplantationofGFPovarytissue（卵巢细胞移植）哺乳动物成年卵巢不含有生殖干细胞(Germline

Stem

Cells:

GSC)JonathanL.TillyGroup？Nature2004;428:145–50.GosdenRG.HumanitRepUpdate2004;10:193–5.GreenfeldC,FlawsJA.Bioessays2004;26:829–32.ByskovAG,FaddyMJ,LemmenJG,AndersenCY.Eggsforever?Differentiation2005;73:438–46.Bristol-GouldSK,KreegerPK,SelkirkCG,KilenSM,MayoKE,SheaLD,etal.DevBiol2006;298:149–54.JohnGB,ShirleyLJ,GallardoTD,CastrillonDG.Reproduction2007;133:855–63.MottaPM,MakabeS.JSubmicroscCytol1986;18:271–90.AlbertiniDF.Reproduction2004;127:513–4.骨髓干细胞可以补充卵巢干细胞（2005）OocyteGenerationinAdultMammalianOvariesbyPutativeGermCellsinBoneMarrowandPeripheralBlood；Cell.122,2005,Pages303–315Folliclenumberrecovered2monthsafterdepleteovarianfolliclesbychemotherapydrug（化疗两个月后卵泡数量会恢复）CD51-positiveprimordialgermcellsexpressgermlinemakersOct4,Mvh,Dazl,andStella,butnotoocytemarkers,Gdf9orZp3.Thesemarkerswerealsofoundinbonemarrow.（早期原始生殖细胞跟骨髓干细胞表达一些相似的基因）MicetreatedwithcyclophosphamideandbusulfantodestroyexistingGSCsthatalsoreceivedbonemarrowtransplantshadareturnofoocyte-containingfollicles.（化疗动物接受骨髓移植后，卵泡会比没有骨髓移植动物恢复的快）PeripheralbloodfromGFPmicetoAtm-deficientmiceresultedGFP-positiveoocytesweredetectableinrecipientsovary（

输血动物可以在其卵巢中看到GFP标记的来自供血动物的卵泡细胞）骨髓干细胞可以补充卵巢干细胞OocyteGenerationinAdultMammalianOvariesbyPutativeGermCellsinBoneMarrowandPeripheralBlood；Cell.122,2005,Pages303–315合体实验2009卵巢生殖干细胞的分离

中国上海交大SchoolofLifeScienceandBiotechnology,ShanghaiJiaoTongUniversity,Shanghai,200240,China.FACSisolationofMVH/DDX4-expressingcellsMVH(mousevasahomologueprotein)DDX4(DEADboxpolypeptide4)ZouK,YuanZ,YangZ,LuoH,SunK,ZhouL,etal.Productionofoffspringfromagermlinestemcelllinederivedfromneonatalovaries.NatCellBiol2009;11:631–6.b:Red,BrdU-positivecells.(c)ArrowsindicateMVH+cells.(e)BrdU(green)andMVH(red)争论在继续。。。。WheredoOogonialstemcellslocate?

1.germinativeepithelium(GE) 2.Ovariancortex卵巢生殖干细胞的位置？Structureoftheovary.Thecorticalregionissurroundedbyasimpleepitheliumclassicallynamedgerminativeepithelium(GE).Underlyingtheovariansurfaceepitheliumisaconnectivetissuelayer,thetunicaalbuginea(TA).Groupsofprimordialfollicles(PF)arepresentintheovarianstroma.ColHEstainx40.Marker蛋白的特异性？？生理意义？？YesNotyetYes卵巢含有体细胞系列的干细胞吗？

Somaticstemcellsintheovary?BasicKnowledgeofStemCells A.Definition

B.Classification

C.CharacterizationStemCellsinTestis A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary

A.Germlinestemcells B.Somaticstemcells4.FutureDirections A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithgermlineandsomaticstemcells?外源性（体细胞系列）的干细胞可以帮助卵巢功能恢复Niche功能？