Mammalian Cell Culture

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1、AnIntroductiontoMammalianCellCulture哺乳类动物细胞培养指南•Applications应用•Stemcells干细胞•the“normal“Cell正常细胞•Differentiation分化•Senescenceand•CellDivision分裂Transformation•Proteinproduction蛋白生产•the“Cancer”Cell肿瘤细胞•TissueEngineering组织工程Whyusemammaliancells?原因•Disadvantages:弊•Necessarybecauseof利–

2、lowgrowthrates生长慢–correctformationofdisulfidebondsinproducedproteins蛋白结–expensivemedia花费贵构正确–requireserumandgrowthfactors需要血清和生长因子–correctglycosylation(conformation,biological–lowspecificproductionactivity,in-vivohalflife)糖rateonbiomass产率低基化正确TypicalApplicationsofMammalianCellCul

3、ture常见应用•CellBiologyResearch细胞生物学研究•ToxicityTesting毒理检验•ProductionofComplexProteins蛋白生产•ProductionofVaccines疫苗生产•TissueandOrganCulture组织和器官培养•mostfrequentproduct:monoclonalantibodiesfordiagnosticsandtherapy诊断和治疗的单抗Glossary-cellculture细胞培养名词•tissueculture组织培养maintainanceorgrowthof

4、tissuefragmentsinvitro•organculture器官培养maintainanceorgrowthoftissues,organsorpartsoforgansinvitro•cellculture细胞培养maintainanceandcultureofcellsinvitro•primarycellline初级细胞西cellsisolatedfromtissue,beforepassaging•secondarycellline次级细胞系isolatedcellsafterfirstpassage•diploidcellline倍体

5、细胞系finiteinvitrolifespanchromosomes:structuralidenticaltothoseofthespeciesfromwhichtheywerederived•continuouscellline传代细胞系indefiniteinvitrolifespan(immortal,established)EukaryoticCellstructure真核细胞结构•Organelles器官•Plasmamembrane膜•3-dimensionalstructure三维结构•Polarity极性Influenceofmicr

6、ofilamentsandtubulionphenotype微丝微管TheNormalCell正常细胞•Whateverthatis?Whatisnormal?ACharacterisation特点•Toexistinanorderly,•growthfactordependence生长因子依developmentallyregulated赖性proliferationdependsonavailabilityoftissue-typespecifictissue,cellsgenerallyhavegrowthfactorssomecommon•inm

7、anycasesgrowthfactorcharacteristics:正常细胞的withdrawalleadstoapoptosis特点包括：•需要生长因子，没有的话会凋亡•contactinhibition接触抑制•somaticcellshavelimited•like-cellscontact:inhibitionofcellproliferativecapacity有限的增殖movementandproliferation•replicativesenescence重复性衰老•quiescentcells(G0phaseofcellcycle)

8、•anchoragedependence锚定依赖性•proliferationr