1、會(huì)計(jì)學(xué)1動(dòng)植物細(xì)胞工程動(dòng)植物細(xì)胞工程Figure4-2The fibroblast.(A)Phase-contrastmicrographoffibroblastsinculture.(B)Drawingsofalivingfibroblastlikecellinthetransparenttailofatadpole,showingthechangesinitsshapeandpositiononsuccessivedays.Notethatwhilefibroblastsflattenoutinculture,theycanhavemorecomplex,process-bearingmo

2、rphologiesintissues.(A,courtesyofDanielZicha;B,redrawnfromE.Clark,Am. J. Anat.13:351-379,1912.)Cells/tissues grown in culture long period of timeBASIC STERILE TECHNIQUEWork environment and surfacePlasticware and glasswareHandling techniquesSterilization of solutions for maintenancegrowth or treatmen

3、tnWORK ENVIRONMENT AND SURFACEnLaminar flow hoodn Relatively enclosed spacen Little traffic flowing past work spacen Confined space can be easily cleaned and maintainedn Must have annual checkup of HEPA filtersn Should have internal outlets for electricity, vacuum and gasn Frequently outfitted with

4、UV lightn Prevent contamination by daily scrub with 70% ethanoln Use closed flask attached to vacuum for spent mediumnSeparate sterile roomn All incoming air circulated through HEPA filtersnPLASTICWARE AND GLASSWAREnPipettes, bottles, flasks, petri dishesnPlasticn- better attachment and cell growth

5、of monolayer cell culturen- more expensive than reusable glassn- suitable for storage at 4CnGlassn- Can withstand temperatures 0C use for maintaining frozen stocksn- Steam sterilize by autoclavingn* relies on steam pressure and high heatn* use fast-dry cycle to dry condensaten* use “wet” cycle for s

6、alt solutions toprevent evaporationn* loosely place caps on bottles生長(zhǎng)因子多為肽激素，有胰島素、表皮生長(zhǎng)因素（EGF）、成纖細(xì)胞生長(zhǎng)因素（fibroblastgfowthfactor，F(xiàn)GF）、血小板來(lái)源增殖因素（plateletderivedgrowthfactor，PDGF）以及生長(zhǎng)激素釋放抑制因子（somatostatinSRIH）DMEM培養(yǎng)基培養(yǎng)基胎牛血清胎牛血清其他成分其他成分錐形瓶逐級(jí)錐形瓶逐級(jí)放大培養(yǎng)放大培養(yǎng)加堿（碳酸加堿（碳酸氫鈉）氫鈉）加糖（葡萄加糖（葡萄糖）糖）灌注培養(yǎng)液灌注培養(yǎng)液微載體微載體5L種子罐培

7、種子罐培養(yǎng)養(yǎng)培養(yǎng)液培養(yǎng)液50L生生物物反反應(yīng)應(yīng)器器接種狂犬接種狂犬病毒病毒出液口出液口收獲病毒收獲病毒組織工程（TissueEngineering）是近年來(lái)正在興起的一門新興學(xué)科，組織工程一詞最早是由美國(guó)國(guó)家科學(xué)基金會(huì)1987年正式提出和確定的。它是應(yīng)用生命科學(xué)和工程學(xué)的原理與技術(shù)，在正確認(rèn)識(shí)哺乳動(dòng)物的正常及病理兩種狀態(tài)下結(jié)構(gòu)與功能關(guān)系的基礎(chǔ)上，研究、開發(fā)用于修復(fù)、維護(hù)、促進(jìn)人體各種組織或器官損傷后的功能和形態(tài)生物替代物的科學(xué)。組織工程的核心就是建立細(xì)胞與生物材料的三維空間復(fù)合體，即具有生命力的活體組織，用以對(duì)病損組織進(jìn)行形態(tài)、結(jié)構(gòu)和功能的重建并達(dá)到永久性替代。共基本原理和方法是將體外培養(yǎng)擴(kuò)增

8、的正常組織細(xì)胞，吸附于一種生物相容性良好并可被機(jī)體吸收的生物材料上形成復(fù)合物，將細(xì)胞-生物材料復(fù)合物植入機(jī)體組織、器官的病損病分，細(xì)胞在生物材料逐漸被機(jī)體降解吸收的過程中形成新的在形態(tài)和功能方面與相應(yīng)器官、組織相一致的組織，而達(dá)到修復(fù)創(chuàng)傷和重建功能的目的。生物相容性好、可被人體降解吸收的組織工程支架材料稱為細(xì)胞外基質(zhì)（ECM），其功能是為細(xì)胞提供生存空間，使細(xì)胞獲足夠的營(yíng)養(yǎng)物質(zhì)，進(jìn)行氣體交換，并使細(xì)胞按預(yù)制形態(tài)的三維支架生長(zhǎng)。在細(xì)胞和生物材料的復(fù)合體植入機(jī)體病損部位后，生物支架被降解吸收，但種植的細(xì)胞繼續(xù)增殖繁殖，形成新的具有原來(lái)特殊功能和形態(tài)的相應(yīng)組織器官。種子細(xì)胞:自體、同種異體、異種組織

9、細(xì)胞等細(xì)胞外基質(zhì)（extracelluarmatrix,ECM）:理想的ECM應(yīng)具有以下特點(diǎn)：生物相容性好，在體內(nèi)不引起炎癥反應(yīng)和毒性反應(yīng)；有可吸收性，能徹底地被自身組織所取代；有可塑性，可塑為任意的三維結(jié)構(gòu)，植入后在體內(nèi)仍可保持特定形狀；表面化學(xué)特性和表面微結(jié)構(gòu)利于細(xì)胞的粘附和生長(zhǎng)；降解速率可根據(jù)不同細(xì)胞的組織再生速率而進(jìn)行調(diào)整。天然:膠原人工:聚乳酸（polylacticacidPLA）、聚羥基乙酸（polyglycolicacidPGA）、兩者的共聚物（PGA-PLA）、聚-羥基丁酯（PHB）；聚乳酸-已內(nèi)酯的共聚物（PLC）、聚原酸酯、聚磷本酯、聚酸酐等。組織工程臨床應(yīng)用:組織工程中臨

10、床應(yīng)用是在組織構(gòu)建完成了動(dòng)物試驗(yàn)之后，在人體上的應(yīng)用，這也是組織工程的最后一步。目前，組織工程的研究只有活性皮膚達(dá)到了這一步。Figure22-1Mammalian skin.(A)Schematicdiagramsshowingthecellulararchitectureofthickskin.(B)Photographofacross-sectionthroughthesoleofahumanfoot,stainedwithhematoxylinandeosin.Theskincanbeviewedasalargeorgancomposedoftwomaintissues:epithel

11、ialtissue(theepidermis),whichliesoutermost,andconnectivetissue,whichconsistsofthetoughdermis(fromwhichleatherismade)andtheunderlyingfattyhypodermis.Eachtissueiscomposedofavarietyofcelltypes.Thedermisandhypodermisarerichlysuppliedwithbloodvesselsandnerves.Somenervefibersextendalsointotheepidermis.Fig

12、ure22-19Cross-section of mammalian epidermis.(A)Schematicdiagram.(B)Photomicrographofasectionthroughthesoleofthefoot(hematoxylinandVanGiesonstain).Thegranular cellsbetweenthepricklecellsandtheflattenedsquamesareinthepenultimatestagesofkeratinization;theyappeargranularbecausetheycontaindarklystaining

13、aggregatesofamaterialcalledkeratohyalin,whichisthoughttobeinvolvedintheintracellularcompactionandcross-linkingofthekeratin.Keratohyalinconsistsmainlyofaproteinknownasfilaggrin.Inadditiontothecellsdestinedforkeratinization,thedeeplayersoftheepidermisincludesmallnumbersofcellsofdifferentcharacter(nots

14、hownhere)includingmacrophagelikeLangerhanscells,derivedfrombonemarrow;melanocytes,derivedfromtheneuralcrest;andMerkel cells,whichareassociatedwithnerveendingsintheepidermis.SeealsoFigure22-1.Figure22-21The columnar organization of squames in the epidermal layer of thin skin.Thestructureisrevealedbys

15、wellingthekeratinizedsquamesinasolutioncontainingsodiumhydroxide.Thistypeoforganizationoccursonlywheretheepidermisisthin.Somestudiessuggestthateachsuchcolumnisaproliferativeunit,correspondingtoasinglestemcellamongthe10-12basalcellsonwhichthecolumnrests.Figure22-22An immortal stem cell.Eachself-renew

16、ingpatchofepidermismustcontainineachcellgenerationatleastoneimmortalstemcell,whosedescendantswillstillbepresentinthepatchinthedistantfuture.Thearrowsindicatelinesofdescent.Animmortalstemcellisshownhereoccupyingthesamepositionineachcellgeneration.Otherbasalcellsmightbebornchemicallydifferentinawaytha

17、tcommitsthemtoleavethebasallayeranddifferentiate;ortheytoomightbestemcells,equivalenttotheimmortalstemcellincharacterandmortalonlyinthesensethattheirprogenyhappensubsequentlytobejostledoutofthebasallayerandshedfromtheskin.Figure22-37Myoblast fusion in culture.Thephase-contrastmicrographsshowhowthece

18、llswillproliferate,lineup,andfusetoformmultinucleatemusclecells.(C)isathighermagnification,showingthecross-striationsthatarejustbeginningtobevisibleasthecontractileapparatusdevelops(red arrow)andtheaccumulationsofmanynucleiwithinasinglecell(green arrows).(CourtesyofRosalindZalin.)Figure22-39Autoradi

19、ograph of a single multinucleate muscle cell with associated satellite cells.ThefiberhasbeenisolatedfromanadultratandtransferredintoculturemediumcontainingH-thymidineplusanextractfromdamagedmusclethatstimulatesthesatellitecellstodivide.Thedividingsatellitecells(arrows)havebecomeradioactivelylabeled(

20、silvergrainsvisibleasblack dots);themusclecellnucleiareunabletoproliferateandremainunlabeled.(FromR.Bischoff,Dev. Biol.115:140-147,1986.)Figure21-99A typical neuron of a vertebrate.Thearrowsindicatethedirectioninwhichsignalsareconveyed.Theneuronshownisfromtheretinaofamonkey.Thelongestandlargestneuro

21、nsinahumanextendforabout1millionmmandhaveanaxondiameterof15mm.Figure21-100The complex organization of nerve cell connections.Thissemischematicdrawingdepictsasectionthroughasmallpartofamammalianbraintheolfactorybulbofadog,stainedbytheGolgitechnique.Theblackobjectsareneurons;thethinlinesareaxonsandden

22、drites,throughwhichthevarioussetsofneuronsareinterconnectedaccordingtopreciserules.Figure21-101The three phases of neural development.Figure21-102Diagram of an early (2 1/2-day) chick embryo, showing the origins of the nervous system.Theneuraltube(light green)hasalreadyclosed,exceptatthetailend,andl

23、iesinternally,beneaththeectoderm,ofwhichitwasoriginallyapart.Theneuralcrest(red)liesdorsallybeneaththeectoderm,inorabovetheroofoftheneuraltube.Inaddition,thickenings,orplacodes(dark green),intheectodermoftheheadgiverisetosomeofthesensorytransducercellsandneuronsofthatregion,includingthoseoftheearand

24、thenose.Thecellsoftheretinaoftheeye,bycontrast,originateaspartoftheneuraltube.Figure21-103Formation of the neural tube.Thescanningelectronmicrographshowsacross-sectionthroughthetrunkofa2-daychickembryo.Theneuraltubeisabouttocloseandpinchofffromtheectoderm;atthisstageitconsists(inthechick)ofanepithel

25、iumthatisonlyonecellthick.Figure21-109NGF effects on neurite outgrowth.Dark-fieldphotomicrographsofasympatheticganglionculturedfor48hourswith(above)orwithout(below)NGF.NeuritesgrowoutfromthesympatheticneuronsonlyifNGFispresentinthemedium.EachculturealsocontainsSchwann(glial)cellsthathavemigratedouto

26、ftheganglion;thesearenotaffectedbyNGF.NeuronalsurvivalandmaintenanceofgrowthconesforneuriteextensionrepresenttwodistincteffectsofNGF.Theeffectongrowthconesislocal,direct,rapid,andindependentofcommunicationwiththecellbody;whenNGFisremoved,thedeprivedgrowthconeshalttheirmovementswithinaminuteortwo.The

27、effectofNGFoncellsurvivalislessimmediateandisassociatedwithuptakeofNGFbyendocytosisanditsintracellulartransportbacktothecellbody.Figure22-43The growth of cartilage.Thetissueexpandsasthechondrocytesdivideandmakemorematrix.Thefreshlysynthesizedmatrixwithwhicheachcellsurroundsitselfisshadeddark green.

28、Cartilagemayalsogrowbyrecruitingfibroblastsfromthesurroundingtissueandconvertingthemintochondrocytes.Figure22-44Deposition of bone matrix by osteoblasts.Osteoblastsliningthesurfaceofbonesecretetheorganicmatrixofbone(osteoid)andareconvertedintoosteocytesastheybecomeembeddedinthismatrix.Thematrixcalci

29、fiessoonafterithasbeendeposited.Theosteoblaststhemselvesarethoughttoderivefromosteogenicstemcellsthatarecloselyrelatedtofibroblasts.Figure22-45An osteoclast shown in cross-section.Thisgiant,multinucleatedcellerodesbonematrix.Theruffledborderisasiteofsecretionofacids(todissolvetheboneminerals)andhydr

30、olases(todigesttheorganiccomponentsofthematrix).Osteoclastsvaryinshape,aremotile,andoftensendoutprocessestoresorbboneatmultiplesites.Theydevelopfrommonocytesandcanbeviewedasspecializedmacrophages.(FromR.V.Krstic,UltrastructureoftheMammalianCell:AnAtlas.Berlin:Springer,1979.)Figure22-46The remodeling

31、 of compact bone.Osteoclastsactingtogetherinasmallgroupexcavateatunnelthroughtheoldbone,advancingatarateofabout50mmperday.Osteoblastsenterthetunnelbehindthem,lineitswalls,andbegintoformnewbone,depositinglayersofmatrixatarateof1or2mmperday.Atthesametimeacapillarysproutsdownthecenterofthetunnel.Thetun

32、nelwilleventuallybecomefilledwithconcentriclayersofnewbone,withonlyanarrowcentralcanalremaining.Eachsuchcanal,besidesprovidingarouteofaccessforosteoclastsandosteoblasts,containsoneormorebloodvesselsbringingthenutrientsthebonecellsmusthavetosurvive.Typically,about5-10%oftheboneinahealthyadultmammalis

33、replacedinthiswayeachyear.Figure22-47Transverse section through a compact outer portion of a long bone.Themicrographshowstheoutlinesoftunnelsformedbyosteoclastsandthenfilledinbyosteoblastsduringsuccessiveroundsofboneremodeling.Thesectionhasbeenpreparedbygrinding;thehardmatrixhasbeenpreservedbutnotth

34、ecells.Lacunaeandcanaliculithatwereoccupiedbyosteocytesareclearlyvisible,however.Thealternatingbrightanddarkconcentricringscorrespondtoanalternatingorientationofthecollagenfibersinthesuccessivelayersofbonematrixlaiddownbytheosteoblaststhatlinedthewallofthecanalduringlife.(Thispatternisrevealedhereby

35、viewingthespecimenbetweenpartlycrossedPolaroidfilters.)Notehowoldersystemsofconcentriclayersofbonehavebeenpartlycutthroughandreplacedbynewersystems.Figure22-48The development of a long bone.Longbones,suchasthefemurorthehumerus,developfromaminiaturecartilagemodel.Uncalcifiedcartilageisshowningreen,ca

36、lcifiedcartilageinblack,boneinbrown,andbloodvesselsinred.Thecartilageisnotconvertedtobonebutisgraduallyreplacedbyitthroughtheactionofosteoclastsandosteoblasts,whichinvadethecartilageinassociationwithbloodvessels.Osteoclastserodecartilageandbonematrix,whileosteoblastssecretebonematrix.Theprocessofoss

37、ificationbeginsintheembryoandisnotcompleteduntiltheendofpuberty.Theresultingboneconsistsofathick-walledhollowcylinderofcompactboneenclosingalargecentralcavityoccupiedbythebonemarrow.Notethatnotallbonesdevelopinthisway.Themembrane bonesoftheskull,forexample,areformeddirectlyasbonyplates,notfromaprior

38、cartilagemodel.Figure22-49Remodeling of a long bone in the leg after a fracture that has healed out of true.Thedeformityintherecentlyhealedboneexposesittoabnormalstresses.Wherethecompressiveforcesareincreased,therateofbonedepositionisincreasedrelativetotherateoferosion;wheretheforcesaredecreased,the

39、rateofdepositionisdecreasedrelativetotherateoferosion.Inthiswaytheboneisgraduallyremodeledbacktoitsnormalform.Figure22-13Angiogenesis.Anewbloodcapillaryformsbythesproutingofanendothelialcellfromthewallofanexistingsmallvessel.Thisschematicdiagramisbasedonobservationsofcellsinthetransparenttailofalivi

40、ngtadpole.(AfterC.C.Speidel,Am. J. Anat.52:1-79,1933.)Figure22-14Capillary formation in vitro.Endothelialcellsinculturespontaneouslydevelopinternalvacuolesthatjoinup,givingrisetoanetworkofcapillarytubes.Photographs(A)and(B)showsuccessivestagesintheprocess;thearrowin(A)indicatesavacuoleforminginitial

41、lyinasingleendothelialcell.Theculturesaresetupfromsmallpatchesoftwotofourendothelialcellstakenfromshortsegmentsofcapillary.Thesecellswillsettleonthesurfaceofacollagen-coatedculturedishandformasmallflattenedcolonythatenlargesgraduallyasthecellsproliferate.Thecolonyspreadsacrossthedish,andeventually,a

42、fterabout20days,capillarytubesbegintoforminthecentralregions.Oncetubeformationhasstarted,branchessoonappear,andafter5to10moredaysanextensivenetworkoftubesisvisible,asseenin(B).Figure22-25White blood cells.(A-D)Electronmicrographsshowing,respectively,aneutrophil,abasophil,aneosinophil,andamonocyte.El

43、ectronmicrographsoflymphocytesareshowninFigure23-4.Eachofthecelltypesshownherehasadifferentfunction,whichisreflectedinthedistinctivetypesofsecretorygranulesandlysosomesitcontains.Thereisonlyonenucleuspercell,butithasanirregularlobedshape,andin(B),(C),and(D)theconnectionsbetweenthelobesareoutofthepla

44、neofsection.(E)LightmicrographofabloodsmearstainedwiththeRomanowskystain,whichcolorsthewhitebloodcellsstrongly.(A-D,courtesyofDorothyBainton;E,courtesyofDavidMason.)Figure22-27Bone marrow.(A)Lightmicrographofastainedsection.Thelargeemptyspacescorrespondtofatcells,whosefattycontentshavebeendissolveda

45、wayduringspecimenpreparation.Thegiantcellwithalobednucleusisamegakaryocyte.(B)Low-magnificationelectronmicrograph.Thistissueisthemainsourceofnewbloodcells(exceptforTlymphocytes,whichareproducedinthethymus).Notethattheimmaturebloodcellsofaparticulartypetendtoclusterinfamilygroups.(A,courtesyofDavidMa

46、son;B,fromJ.A.G.Rhodin,Histology:ATextandAtlas.NewYork:OxfordUniversityPress,1974.)Figure22-30A tentative scheme of hemopoiesis.Thepluripotentstemcellnormallydividesinfrequentlytogenerateeithermorepluripotentstemcells(self-renewal)orcommitted progenitor cells(labeledCFC=colony-formingcells),whichare

47、irreversiblydeterminedtoproduceonlyoneorafewtypesofbloodcells.Theprogenitorcellsarestimulatedtoproliferatebyspecificgrowthfactorsbutprogressivelylosetheircapacityfordivisionanddevelopintoterminallydifferentiatedbloodcells,whichusuallyliveforonlyafewdaysorweeks.Inadultmammalsallofthecellsshowndevelop

48、mainlyinthebonemarrowexceptforTlymphocytes,whichdevelopinthethymus,andmacrophagesandosteoclasts,whichdevelopfrombloodmonocytes.ThemostcontroversialpartoftheschemeiswheretheprecursorsforTandBlymphocytesfitintothescheme.Thedashedlinesreflectthisuncertainty.Thepluripotentstemcellsalsogiverisetovarioust

49、ypesoftissuecellsnotshowninthisscheme,suchasNKcells,mastcells,andavarietyofclassesofantigen-presentingcells(discussedinChapter23),butthepathwaysbywhichthesecellsdevelopareuncertain.Figure22-32The development of red blood cells.ThedrawingshowstherelationshipbetweentheBFC-E,theCFC-E,andthematureerythr

50、ocyte.BFC-EsandCFC-Esarebothcommittederythroidprogenitorcells.BFC-EsrespondtothefactorIL-3butnottoerythropoietin,whereasCFC-Esrespondtoerythropoietin.Theseriesofcelldivisionsthatoccurinthislineageundertheinfluenceoferythropoietinprovidesapowerfulmeansofcontrollingtheproductionoferythrocyteswithoutup

51、settingtheproductionofothertypesofbloodcells.Figure22-16Renewal of the gut lining.(A)Thepatternofcellturnoverandtheproliferationofstemcellsintheepitheliumthatformstheliningofthesmallintestine.Thenondividingdifferentiatedcellsatthebaseofthecryptsalsohaveafinitelifetime,terminatedbyprogrammedcelldeath

52、,andarecontinuallyreplacedbyprogenyofthestemcells.(B)Photographofasectionofpartoftheliningofthesmallintestine,showingthevilliandcrypts.Notehowmucus-secretinggobletcells(stainedred)areinterspersedamongtheabsorptivebrush-bordercellsintheepitheliumofthevilli.SeeFigure22-9forthestructureofthesecells.Mil

53、estones in Stem Cell ResearchMouse ES cells (Evens MJ and Kaufman, 1981)Cloning of Dolly (Wilmut, 1997)Establishing human ES cells (Thomson, 1998Trans-differentiation (Goodale; Verfaillie, 2003)Human ES cells by SCNT (Huang, 2005)Generation of iPS cells (Yamanaka, 2006）The definition of a stem cell.

54、Eachdaughterproducedwhenastemcelldividescaneitherremainastemcellorgoontobecometerminallydifferentiated.Figure21-29The early stages of mouse development.Figure21-30Scanning electron micrographs of the early mouse embryo.Thezonapellucidahasbeenremoved.(A)Two-cellstage.(B)Four-cellstage(apolarbodyisvis

55、ibleinadditiontothefourblastomeresseeFigure20-16).(C)Eight-to-sixteen-cellmorulacompactionoccurring.(D)Blastocyst.Figure21-31A procedure for creating a chimeric mouse.Twomorulaeofdifferentgenotypesarecombined.Figure21-32Making a chimeric mouse with ES or teratocarcinoma stem cells.Theexperimentshows

56、thatthestemcellscancombinewiththecellsofanormalblastocysttoformahealthychimericmouse.Shinya YamanakaCell 2007Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined FactorsOct4/Sox2/Myc/Klf4干細(xì)胞基礎(chǔ)研究的重大問題自我復(fù)制（Self-renewal）和多分化潛能(Pluripotency)的維持胚胎干細(xì)胞向各種組織的功能細(xì)胞的分化調(diào)控干細(xì)胞基因組重編程的機(jī)制(SCNT

57、，iPS cells)腫瘤干細(xì)胞腫瘤細(xì)胞是否由干細(xì)胞變異而來(lái)？腫瘤對(duì)治療的耐受是否由于存在腫瘤干細(xì)胞6.1 單克隆抗體單克隆抗體 1975 1975年英國(guó)科學(xué)家年英國(guó)科學(xué)家Milstein和和Kohler將產(chǎn)生抗體將產(chǎn)生抗體 的淋巴細(xì)胞同腫瘤細(xì)胞融合，成功建立了單克隆抗的淋巴細(xì)胞同腫瘤細(xì)胞融合，成功建立了單克隆抗 體技術(shù)，而獲得體技術(shù)，而獲得1984年諾貝爾醫(yī)學(xué)和生理學(xué)獎(jiǎng)。年諾貝爾醫(yī)學(xué)和生理學(xué)獎(jiǎng)。 每個(gè)每個(gè)B淋巴細(xì)胞僅專一地產(chǎn)生、分泌一種針對(duì)某淋巴細(xì)胞僅專一地產(chǎn)生、分泌一種針對(duì)某 種抗原決定簇的特異性抗體，而腫瘤細(xì)胞可以無(wú)限種抗原決定簇的特異性抗體，而腫瘤細(xì)胞可以無(wú)限 增殖，因此雜交瘤細(xì)胞可在

58、體外培養(yǎng)或移植到體內(nèi)增殖，因此雜交瘤細(xì)胞可在體外培養(yǎng)或移植到體內(nèi) 條件下分泌大量單克隆抗體。條件下分泌大量單克隆抗體。 單克隆抗體技術(shù)的最主要優(yōu)點(diǎn)是可以用不純的抗單克隆抗體技術(shù)的最主要優(yōu)點(diǎn)是可以用不純的抗 原分子大量制備純一的單克隆抗體。原分子大量制備純一的單克隆抗體。雜交瘤細(xì)胞產(chǎn)生雜交瘤細(xì)胞產(chǎn)生 單克隆抗體示意圖單克隆抗體示意圖6.2 動(dòng)物克隆動(dòng)物克隆 生物繁殖后代通常是以精、卵細(xì)胞結(jié)合的有性生殖生物繁殖后代通常是以精、卵細(xì)胞結(jié)合的有性生殖方式進(jìn)行。通過營(yíng)養(yǎng)體細(xì)胞繁殖個(gè)體的方法稱為無(wú)性繁方式進(jìn)行。通過營(yíng)養(yǎng)體細(xì)胞繁殖個(gè)體的方法稱為無(wú)性繁殖?？寺∈侵鸽x體條件下的無(wú)性繁殖。殖?？寺∈侵鸽x體條件下的

59、無(wú)性繁殖。1981年年Illmenses 率先報(bào)告用小鼠幼胚細(xì)胞核克隆出正常小鼠。隨后，率先報(bào)告用小鼠幼胚細(xì)胞核克隆出正常小鼠。隨后，1984年年Willadsen用未成熟羊胚細(xì)胞核克隆出一頭羊。用未成熟羊胚細(xì)胞核克隆出一頭羊。 英國(guó)英國(guó)PPI生物技術(shù)的羅斯林（生物技術(shù)的羅斯林（Roslin）研究所的維爾）研究所的維爾穆特穆特(Wilmut)博士博士1997年年2月月27日在世界著名權(quán)威雜志日在世界著名權(quán)威雜志Nature宣布的用乳腺細(xì)胞的細(xì)胞核克隆出一只綿羊宣布的用乳腺細(xì)胞的細(xì)胞核克隆出一只綿羊“ 多莉多莉 (Dolly) ”的消息。的消息?！岸嗬蚨嗬颉钡恼Q生，既說(shuō)明了體細(xì)的誕生，既說(shuō)明了體

60、細(xì)胞核的遺傳全能性，也翻開了人類以體細(xì)胞核競(jìng)相克隆胞核的遺傳全能性，也翻開了人類以體細(xì)胞核競(jìng)相克隆哺乳動(dòng)物的新篇章。此項(xiàng)技術(shù)因而榮登美國(guó)哺乳動(dòng)物的新篇章。此項(xiàng)技術(shù)因而榮登美國(guó)Science周刊評(píng)出的周刊評(píng)出的1997 年十大科學(xué)發(fā)現(xiàn)年十大科學(xué)發(fā)現(xiàn) 的榜首。的榜首。 綿羊綿羊B乳腺細(xì)胞核乳腺細(xì)胞核易核卵易核卵融合卵融合卵綿羊綿羊C子宮子宮多莉多莉植入植入植入植入生產(chǎn)生產(chǎn)克克隆隆多多莉莉羊羊示示意意圖圖取出取出未受精卵未受精卵去核去核電激電激體外胚胎發(fā)育體外胚胎發(fā)育綿羊綿羊A多莉多莉克隆猴克隆猴 (1)遺傳素質(zhì)完全一致的克隆動(dòng)物將更有利于開展對(duì)動(dòng)遺傳素質(zhì)完全一致的克隆動(dòng)物將更有利于開展對(duì)動(dòng)物物(人