BasicKnowledgesaboutStemCells(干細胞基本知識) A.Definition

B.Classification

C.Characterization2.StemCellsinTestis(睪丸干細胞) A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary(卵巢干細胞）

A.Germlinestemcells

B.Somaticstemcells4.FutureDirections(未來方向) A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本課內容Stemcellsarethecellsthatcandivideessentiallywithoutlimittoformothercells.(無限分裂并形成其他細胞的細胞）Self-renewaltomaintainthemselves

(自我增殖)2)Differentiationtoformspecializedcells

(分化成子代細胞）Whatarestemcells?全能干細胞

(Totipotent

stemcells):Cells

canformallthecelltypesinabody,plustheextraembryonic,orplacental,cells.Embryoniccellswithinthefirstcoupleofcelldivisionsafterfertilizationaretheonlycellsthataretotipotent

(包括胎盤).多能干細胞

(Pluripotent

stemcells):Cellscangiverisetoallofthecelltypesthatmakeupthebody;embryonicstemcellsareconsideredpluripotent

(不包括胎盤).亞多能干細胞

(Multipotent

stemcells):Cellscandevelopintomorethanonecelltype,butaremorelimitedthanpluripotentcells;adultstemcellsandcordbloodstemcellsareconsideredmultipotent.單能干細胞

(Unipotent

stemcells):Cellreferstoacellthatcandifferentiatealongonlyonelineage.干細胞的多能性Totipotent滋養層囊胚Multipotentstemcells（亞多能干細胞）骨髓祖細胞淋巴祖細胞

1.Embryonicstemcells(胚胎干細胞）

Generatedfromcellsofapreimplantation-stageembryo

Totipotent

Pluripotent

2.“Somatic”or“adult”stemcells(成體干細胞)

Generatedfromdevelopingorwell-developedoradultorgans

Multipotent

Unipotent3.Inducedpluripotentstemcells(iPSCs)

(誘導多潛能干細胞) Generatedfromtransductionof4transcriptionalfactors(Oct4 (Pou5f1),Sox2,cMyc,andKlf4)

PluripotentClassification(分類）干細胞小巢及干細胞對稱性和非對稱性分裂干細胞小巢Stem-cellnicheThespecificanatomiclocationwherestemcellslive,whichcontainsextracelluarmatrixandregulatorycells.Theniche(home)playsimportantrolesindeterminingstemcells’fate.Theword'niche'oftenbereferedtoasstem-cellmicroenvironmentinvivoorinvitro.小巢是指干細胞“生活”的位置，其中包含細胞外基質和調控細胞。小巢（家）在調控干細胞的活動及確定干細胞的命運中起著重要的作用?！靶〕病币辉~通常被泛指體內或體外干細胞生存的微環境?！皻w巢”是指干細胞從身體的其它位置，向其小巢回歸的過程。BasicKnowledgesaboutStemCells A.Definition

B.Classification

C.CharacterizationStemCellsinTestis A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary

A.Germlinestemcells B.Somaticstemcells4.FutureDirections A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本課內容SPERMATOGENESIS:/file/view/spermatogenesis.jpg/298006750/800x446/spermatogenesis.jpg曲細精管間質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簡單模型Thefragmentationmodelforspermatogonialmultiplicationandstemcellrenewal.DirkG.deRooijDevelopment2017;144:3022-3030?2017.PublishedbyTheCompanyofBiologistsLtd碎片模型ThehierarchicalAsmodel.DirkG.deRooijDevelopment2017;144:3022-3030?2017.PublishedbyTheCompanyofBiologistsLtd分級模型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(簡單模型);Fragmentationmodel(碎片模型);Hierarchicalmodel(分級模型)。TesticularStemCellsforSomaticcells

Sertolicells?(notyet)

Myiodcells?(notyet)

Leydigcells?Yes(PeritubularandPerivescularlocations)CD90綠色--管周細胞(smactin)紅色—間質細胞(P450scc)

培養4周后，在管周myoid細胞的外表面，出現間質細胞BasicKnowledgesaboutStemCells A.Definition

B.Classification

C.CharacterizationStemCellsinTestis A.Germlinestemcells B.Somaticstemcells3.StemCellsinOvary

A.Germlinestemcells B.Somaticstemcells4.FutureDirections A.Germlinestemcells-physiologicalrelevant? B.Assistantreproductionwithinducedoocytes?本課內容植物根尖生長(干細胞功能及頂尖生長）低等動物的卵巢含有生殖干細胞Celegans線蟲（含生殖干細胞）Drosophila果蠅（含生殖干細胞）Medaka日本淡水魚（含生殖干細胞）哺乳動物卵巢含有生殖干細胞嗎？傳統的觀點：卵巢不含生殖干細胞Eggnumberislimitedandcouldbedepletedbyageof50JonathanL.Tilly2004哺乳動物成年卵巢含有生殖干細胞(Germline

Stem

Cells:

GSC)

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

immature

follicles

are

atretic,whichcouldresultindepletionoffolliclesmuchearlier)

（卵泡萎縮速度太快）GermcellmarkersVasahomologue(MVH,aks

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

epithelium).

BrdU

incorporation

of

MVH-positive

cells（卵泡外表達生殖細胞基因的細胞）Germ

cell

toxicant

(busulfan)onlyaffectedfolliclein2weeks,suggestingitaffectedGSC（毒物影響卵泡的速度太慢）OccurrenceofGFP-expressingoocytessurroundedbynon-GFPovariescellsaftertransplantationofGFPovarytissue（卵巢細胞移植）哺乳動物成年卵巢不含有生殖干細胞(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.骨髓干細胞可以補充卵巢干細胞（2005）OocyteGenerationinAdultMammalianOvariesbyPutativeGermCellsinBoneMarrowandPeripheralBlood；Cell.122,2005,Pages303–315Folliclenumberrecovered2monthsafterdepleteovarianfolliclesbychemotherapydrug（化療兩個月后卵泡數量會恢復）CD51-positiveprimordialgermcellsexpressgermlinemakersOct4,Mvh,Dazl,andStella,butnotoocytemarkers,Gdf9orZp3.Thesemarkerswerealsofoundinbonemarrow.（早期原始生殖細胞跟骨髓干細胞表達一些相似的基因）MicetreatedwithcyclophosphamideandbusulfantodestroyexistingGSCsthatalsoreceivedbonemarrowtransplantshadareturnofoocyte-containingfollicles.（化療動物接受骨髓移植后，卵泡會比沒有骨髓移植動物恢復的快）PeripheralbloodfromGFPmicetoAtm-deficientmiceresultedGFP-positiveoocytesweredetectableinrecipientsovary（

輸血動物可以在其卵巢中看到GFP標記的來自供血動物的卵泡細胞）骨髓干細胞可以補充卵巢干細胞OocyteGenerationinAdultMammalianOvariesbyPutativeGermCellsinBoneMarrowandPeripheralBlood；Cell.122,2005,Pages303–315合體實驗2009卵巢生殖干細胞的分離

中國上海交大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)爭論在繼續。。。。WheredoOogonialstemcellslocate?

1.germinativeepithelium(GE) 2.Ovariancortex卵巢生殖干細胞的位置？Structureoftheovary.Thecorticalregionissurroundedbyasimpleepitheliumclassicallynamedgerminativeepithelium(GE).Underlyingtheovariansurfaceepitheliumisaconnectivetissuelayer,thetunicaalbuginea(TA).Groupsofprimordialfollicles(PF)arepresentintheovarianstroma.ColHEstainx40.Marker蛋白的特異性？？生理意義？？YesNotyetYes卵巢含有體細胞系列的干細胞嗎？

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（體細胞系列）的干細胞可以幫助卵巢功能恢復Niche功能？