《动植物细胞工程》PPT课件

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1、4、动物细胞与组织培养4.1动物细胞特点无细胞壁倍增时间长，生长缓慢需氧量少，对搅拌敏感聚集体形成原代细胞培养50代即开始退化4.2动物细胞培养定义动物细胞与组织培养是从动物体内取出细胞或者组织，模拟体内的生理环境，在无菌、适温和丰富的营养条件下，使离体细胞或者组织生存、生长并维持结构和功能的一门技术。是动物细胞工程的基础。类型1.细胞培养；2.组织培养；3器官培养4.3发展历史1885年WRoux鸡胚神经板1907，美国的哈里森使用盖玻片悬滴培养蛙胚神经组织细胞ACarrel无菌技术1951WEarle培养基。。。4.4生长特

2、性贴附生长型如人胚肺细胞，Hela细胞，巨噬细胞，神经细胞上皮样、成纤维样、无定形单层、汇合悬浮生长型细胞如血液白细胞，淋巴组织细胞等Figure 4-2Thefibroblast.(A)Phase-contrastmicrographoffibroblastsinculture.(B)Drawingsofalivingfibroblastlikecellinthetransparenttailofatadpole,showingthechangesinitsshapeandpositiononsuccessivedays.No

3、tethatwhilefibroblastsflattenoutinculture,theycanhavemorecomplex,process-bearingmorphologiesintissues.(A,courtesyofDanielZicha;B,redrawnfromE.Clark,Am.J.Anat.13:351-379,1912.)常用术语Cell,tissue&organculture贴壁依赖性、汇合原/初代培养Primaryculture传代细胞系（有限、无限/连续）[已建成的]二倍体、转化细胞株（有限、无限

4、/连续）克隆接触抑制4.5体外培养细胞基本技术体外培养特点体外培养工具体外培养条件体外细胞生长增殖过程培养方法大规模培养技术4.5.1体外培养特点营养条件苛刻适应性差,敏感培养时间长，易污染Animalcellsaremoredifficulttoculturethanmicroorganismsbecausetheyrequiremanymorenutrientsandtypicallygrowonlywhenattachedtospeciallycoatedsurfaces.Despitethesedifficulties,

5、varioustypesofanimalcells,includingbothundifferentiatedanddifferentiatedones,canbeculturedsuccessfully.4.5.2体外培养工具空心纤维微球Cells/tissuesgrowninculturelongperiodoftimeBASICSTERILETECHNIQUE•Workenvironmentandsurface•Plasticwareandglassware•Handlingtechniques•Sterilization

6、ofsolutionsformaintenancegrowthortreatmentWORKENVIRONMENTANDSURFACELaminarflowhoodRelativelyenclosedspaceLittletrafficflowingpastworkspaceConfinedspacecanbeeasilycleanedandmaintainedMusthaveannualcheckupofHEPAfiltersShouldhaveinternaloutletsforelectricity,vacuumandga

7、sFrequentlyoutfittedwithUVlightPreventcontaminationbydailyscrubwith70%ethanolUseclosedflaskattachedtovacuumforspentmediumSeparatesterileroomAllincomingaircirculatedthroughHEPAfiltersPLASTICWAREANDGLASSWAREPipettes,bottles,flasks,petridishes•Plastic--betterattachmenta

8、ndcellgrowthofmonolayercellculture--moreexpensivethanreusableglass--suitableforstorageat4ºC•Glass--Canwithstandtemperatures<0ºCusef