細胞與細胞工程.ppt

Cell(JL)phasecontrast.(A)NormalHeLacells;(B)treatedwithstaurosporintoinduceapoptosis;or(C)treatedwiththapsigargintoinduceautophagy(Galluzzietal,2007).(D)Apoptotic,and(E)necroptoticJurkatcells(Degterevetal,2005).(F)Paraptotic293Tcells(Sperandioetal,2000;copyright(2000)NationalAcademyofSciences,USA).(G)Necroticand(H)autophagiciBMKcells(Degenhardtetal,2006;Mathewetal,2007).(I)Ras-expressingU251glioblastomacellsshowingmacropinosomes(J.H.Overmeyer,A.Kaul,E.E.Johnson(3)經(jīng)轉(zhuǎn)基因的克隆哺乳動物，將能為人類提供源源不斷的廉價的藥品、保健品以及較易被人體接受的移植器官；(4)科學家將很快地從目前的同種克隆技術(shù)推進到異種克隆,即借腹懷胎的新領(lǐng)域,這無疑將大大促進對瀕臨滅種的哺乳動物的保護工作。,克隆技術(shù)將對21世紀產(chǎn)生的重大影響（？）,第一節(jié)細胞的特征細胞的基礎(chǔ)知識細胞的生長、分化與死亡第二節(jié)動物細胞工程動物細胞培養(yǎng)單克隆抗體動物克隆干細胞工程,Stemcellsarepowerfultoolsinbiologyandmedicine.Whatcanscientistsdowiththesecellstoexploittheirincrediblepotential?,AdaptedfromPrinciplesofBiologypublishedbyNatureEducation.,JAMESKING-HOLMES/SCIENCEPHOTOLIBRARYAmouseembryoisheldinplaceduringnucleartransfer.Theeggsgeneticmaterialhasbeenremovedandanadultcellnucleusisinjectedinitsplace.,四.干細胞工程,Pluripotent:cantransformintoanyoneofthe220celltypesinthehumanbody.Self-renewal:onestemcellcanalsodividetoproducemillionsmorestemcells.,1.WhatareStemCells?,innearlyallanimals,fromtinywormstomiceandhumans.embryonicstemcellsadultstemcellsinducedpluripotentstem(iPS)cells,2.TypesofStemCells,EmbyonicStemCellsVS.AdultStemCells,StemCells:FlexiblefriendsNature483,S22S26,Inthepastdecade,scientistshavealsolearnedtomakestemcellsfromregularmature,differentiatedcells.Thesecellsarecalledinducedpluripotentstemcellsbecausescientistsforcethemtobecomepluripotentevenaftertheyhavereachedadifferentiatedstate.Byturninguptheexpressionofjustafewgenes,scientistscanforceaskincell,forexample,toretraceitsdevelopmentalpathwaybackwardsallthewaytoaflexiblepluripotentstate.,InducedPluripotentStemCells,3.MedicalApplications,Stemcellscanteachusaboutthebiologyofpluripotentcells,howthehumanbodydevelops,andwhathappensduringdisease.Theyarealsoatoolfordevelopingsaferwaystodesignandtestnewmedications.theyarethebasisforregenerativemedicine,usingstemcellstorepairdamagedtissue.,ThediscoveriesofGurdonandYamanakahaveshownthatspecialisedcellscanturnbackthedevelopmentalclockundercertaincircumstances.Althoughtheirgenomeundergoesmodificationsduringdevelopment,thesemodificationsarenotirreversible.Wehaveobtainedanewviewofthedevelopmentofcellsandorganisms.ResearchduringrecentyearshasshownthatiPScellscangiverisetoallthedifferentcelltypesofthebody.Thesediscoverieshavealsoprovidednewtoolsforscientistsaroundtheworldandledtoremarkableprogressinmanyareasofmedicine.iPScellscanalsobepreparedfromhumancells.Forinstance,skincellscanbeobtainedfrompatientswithvariousdiseases,reprogrammed,andexaminedinthelaboratorytodeterminehowtheydifferfromcellsofhealthyindividuals.Suchcellsconstituteinvaluabletoolsforunderstandingdiseasemechanismsandsoprovidenewopportunitiestodevelopmedicaltherapies.,Fromsurprisingdiscoverytomedicaluse,皮膚細胞,iPS,神經(jīng)細胞,心肌細胞,肝細胞,精子、卵子,Whyisatherapeuticcloningapproachimportant?,Bygrowingnewbodypartsfromexistingonesfromthesamepersonwecanavoidtransplantrejectionproblems.Byusingcellsre-engineeredor“cloned”fromourselveswecanbecomeourowntransplantsource-similartobankingbloodforourselvesbeforeasurgicaloperation.,PeopleintheUSaffectedbydiseasesthatmaybehelpedbystemcellresearch,ConditionNumberofPersonsAffectedCardiovasculardiseases58MillionAutoimmunediseases30MillionDiabetes16MillionOsteoporosis10MillionCancer8.2MillionAlzheimersdisease4MillionParkinsonsdisease1.5MillionBurns(severe)0.3MillionSpinalcordinjuries0.25MillionBirthdefects150,000(peryear)Total128.4Million-DatafromthePatientsCoalitionforUrgentResearch,Washington,DC(accordingtoPerry,Ref.267).,NATURE|NEWSChinasstem-cellrulesgounheededHealthministrysattemptatregulationhashadlittleeffect.Nature484,149150(12April2012),Itsboomtimeforfirmssellingstem-celltreatments,suchasthosederivedfromumbilicalblood.LIUHAIFENG/XINHUAPRESS/CORBIS,NATURE|NEWSStem-celltherapytakesoffinTexasAboominunprovenproceduresisworryingscientists.Nature483,1314(01March2012),Celltexhoststhelargeststem-cellbankintheUnitedStates.TYLERRUDICK,4.DevelopmentoftheStemCelltechnique,Source:,WherearetheStemCells?,自然胚胎克隆人胚胎-核移植/混種胚胎-核移植體細胞誘導多能干細胞(體細胞轉(zhuǎn)分化),In1868,GermanbiologistErnstHaeckelfirstusedthetermstemcelltodescribeazygote.Researchershavebeenworkingwithmouseembryonicstemcellssince1981.thefieldtookoffin1998,whenresearchersisolatedhumanembryonicstemcellsforthefirsttime.（最早的人類多能干細胞是從體外受精的臨床病例中得來的多余胚泡。這個里程碑式的事件是JamesThomson領(lǐng)導的UniversityofWisconsin,Madison的實驗室在1998年的成果。）,3.1IsolationandcultureofStemCells,3.2ObjectionstoStemCellReasearch,2008紐卡斯爾大學將從人類皮膚細胞中提取的DNA注入來自母牛卵巢的卵子，卵子的所有遺傳物質(zhì)基本上都被去除。,3.3InterspeciesEmbryos,2003上海第二醫(yī)科大學盛慧珍教授研究組將人的體細胞核植入去核的兔卵細胞中。,3.4AdultStemCells,donotraisetheethicalquestionsthatembryonicstemcellsdobecausenoembryosareneededexistinverysmallnumbersoftenburieddeepinthetissuemainlyusedfortwotreatments:bone-marrowtransplantstorebuildapatientsimmunesystem,andskingraftstoreplaceskinoveraburnorotherinjuryotherapplicationsforadultstemcells？usedrugstoactivateapatientsownadultstemcellstoperformthenecessaryrepairs？,Foralongtime,scientiststhoughtthatonceacelldifferentiated,itcouldneverregainitspluripotency.Theyassumedthatitmustbesetinitsidentitybecauseithadsomanygenesturnedonandoffinwaysparticulartothatcelltype.In2006,however,KazutoshiTakahashiandShinyaYamanakaofKyotoUniversityinJapanprovedotherwise.Theyturnedmouseskincellsspecifically,fibroblastsintostemcells.Thesewerethefirstinducedpluripotentstem(iPS)cells.,3.5InducedPluripotentStemCells,Thefirstdirectreprogrammingofadulthumanfibroblasts.IanWilmutScottishCentreforRegenerativeMedicine,UniversityofEdinburghIn2007issuesofCellandScience,Yamanakaandcolleagues(Takahashietal.,2007)andYuandcolleagues(Yuetal.,2007)demonstratethatexpressionoffourspecifictranscriptionfactorsgivesadulthumanfibroblastsmanyofthecharacteristicsofhumanembryonicstemcells.Refinementsofthisprocedurewillmakeitpossibletoproducepluripotenthumancelllineswithoutuseofanembryo.Thereareprofoundscientificandsocialimplicationsofthisresearch.,CellStemCell,2007Dec13;1(6):593-4.,Idecidedafewweeksagonottopursuenucleartransfer-themethodusedtocreateDollythesheep,hesaid,admittingthenewmethodwaseasiertoacceptsocially.,十年科學進展1.TheDarkGenome：基因組中的“暗物質(zhì)”3.AncientDNA：古生物的DNA5.ReprogrammingCells：細胞重編程6.TheMicrobiome：人體內(nèi)微生物組8.Inflammation：炎癥反應(yīng)的陰暗面,Science,Dec.2010,TheNobelPrizeinPhysiologyorMedicine2012.N.21Feb2013/nobel_prizes/medicine/laureates/2012/,TheNobelPrizerecognizestwoscientistswhodiscoveredthatmature,specialisedcellscanbereprogrammedtobecomeimmaturecellscapableofd