组织培养

TissueCultureTechnique(InVitroCulture)I.IntroductionⅡ.TypesofcellcultureⅢ.EquipmentⅣ.CleaningandDisinfectionContentI.Introduction1.Conception2.Developinghistory3.Application1.Conception(1)InvitroculturewholecultureorganculturepartsculturetissueculturecellcultureOrganculture•Themaintenanceorgrowthoforganprimordiaorthewholeorpartsofanorganinvitro,inawaythatmayallowdifferentiationandpreservationofthearchitectureand/orfunction.Tissueculture•Themaintenanceorgrowthoftissueinvitro,inawaythatmayallowdifferentiationandpreservationofthearchitectureand/orfunction.Cellculture•Thistermisusedtodenotethegrowingofcellinvitro,includingthecultureofsinglecell.Incellculture,thecellsarenolongerorganizedintotissues.1.Conception(2)Invivoculturetransplantationstemcell2.Developinghistory1885Roux1907Harrison1910Carrel1950sAseptictechniqueAntibioticsMediumCulturemethods-Microcarrierculture,andsoon.3.Application(1)Biologicproduct(biologics)(2)Virology(3)Stemcellculture(4)Genetics(5)Cytotoxictest(6)ImmunologyⅡ.Typesofcellculture1.Primaryculture2.cellline3.termsofassociatedwithcellculture1.PrimarycultureAculturestartfromcells,tissuesororganstakendirectlyfromorganisms.Primarycell2.CelllineAcelllinearisesfromaprimarycultureatthetimeoffirstsuccessfulsubculture.(1)Finitedcellline(2)infinitedcellline(1)FinitedcelllineAcultureiscapableofonlyalimitednumberofpopulationdoublings,afterwhichthecultureceasesproliferation.Finitedcellline(2)Infinitedcellline(continuouscellculture)Aculturewhichisapparentlycapableofanunlimitednumberofpopulationdoublings,oftenreferredtoasanimmortalcellculture.suchcellsmayormaynotexpressthecharacteristicsofinvitroneoplasticormalignanttransformation.Infinitedcellline3.termsofassociatedwithcellculture(1)cellstrain(2)cellclone(3)passage(1)cellstrainAcellstrainisderivedeitherfromaprimarycultureoracelllinebytheselectionorcloningofcellshavingspecificpropertiesandmarkers.Suchas:speciesmarkers-speciesspecificantigens,isoenzymesandchromosomes.(2)cellcloneInanimalcellcultureterminologyapopulationofcellderivedfromasinglecellbymitoses.(3)passagethetransferortransplantationofcell,withorwithoutdilution,fromoneculturevesseltoanother.Ⅲ.Equipment1.Flaskofliquidnitrogen2.Cultureflasks3.Multiwellplate4.filtersvacuumfilterationsyringe5.invertedmicroscopy6.Refrigerator(lowtemperature)1.Flaskofliquidnitrogen2.Cultureflasks3.Multiwellplate4.1vacuumfilteration4.2syringeⅣ.cleaninganddisinfectionreference1.鄂征.组织培养和分子细胞学技术.1995,北京出版社2.Doyle主编.CellandTissueCultureforMedicalResearch,20003.薛庆善.体外培养的原理与技术.2001,科学出版社4.章静波主编.组织和细胞培养技术.20025.JulioE.Celis主编.细胞生物学实验手册——细胞和组织培养相关技术，病毒，抗体，免疫细胞化学.2006