TissueandCellCultureTechnologyTissueandCellCultureTechnologyGlossarymicropropagation微繁殖技术totipotency全能，全能性fetal胎儿platelet血小板pathologist病理学家colourant着色剂pharmaceutical药品，药物Typesofculture•Tissueculture–First50yearsusedfragmentsoftissueinadishandoutgrowthofcellsfromit•Cellculture(lasthalfof20thcentury)–Cellsdispersedfromtissue,primaryculture,celllineorcellstrainandputinculture–Usesmechanical,enzymaticorchemicaldisaggregation•Organculture–3Dstructureofundisaggregatedtissue•Organotypic(器官型)culture–Mixingcellsofdifferentlineagestoforma3Dstructure.Whatisit?•Tissuecultureisthetermusedfor“theprocessofgrowingcellsartificiallyinthelaboratory”(OSMS.otago.ac.nz/main/bursary)•Tissuecultureinvolvesbothplantandanimalcells•Tissuecultureproducesclones,inwhichallproductcellshavethesamegenotype(unlessaffectedbymutationduringculture)Planttissueculture,alsocalledmicropropagation,isapracticetechniqueinwhichsmalltissuepiecesororgansareremovedfromadonorplantandculturedasepticallyonanutrientmedium.Bymanipulatingthechemicalcompositionofthenutrientmediumandotherenvironmentalparameters,thegrowthanddevelopmentofthetissuesinculturecanbedirectedintodifferentchannels.Whatisneeded?Tissueculture,bothplantandanimalhasseveralcriticalrequirements:•Appropriatetissue(sometissuesculturebetterthanothers)•Asuitablegrowthmediumcontainingenergysourcesandinorganicsaltstosupplycellgrowthneeds.Thiscanbeliquidorsemisolid•Aseptic(sterile)conditions,asmicroorganismsgrowmuchmorequicklythanplantandanimaltissueandcanoverrunacultureWhatisNeeded,II•Growthregulators-inplants,bothauxins&cytokinins.Inanimals,thisisnotaswelldefinedandthegrowthsubstancesareprovidedinserumfromthecelltypesofinterest•FrequentsubculturingtoensureadequatenutritionandtoavoidthebuildupofwastemetabolitesCulturing(micropropagating)PlantTissue-thesteps•Selectionoftheplanttissue(explant)fromahealthyvigorous‘motherplant’-thisisoftentheapicalbud,butcanbeothertissue•Thistissuemustbesterilizedtoremovem...
