生物反应器课件 Different cell properties Select cell culture method

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1、DifferentcellpropertiesSelectcellculturemethodContentIntroductionCellculturespecificationSelectionofbioreactorMicroorganismsBacteria(0.5–2µm)Escherichiacoli(lowerintestine)Yeast(2–5µm)Saccharomycescerevisiae(baker’syeast)Fungi(2–15µm)Penicilliumspec.(Cheesea

2、ndantibiotic)Mammalian(7–20µm)ChineseHamsterOvaryCells(Insuline)Animalcellsvs.microbialcellsPROTEINS-PosttranslationalmodificationsGlycosylation,phosphorylationetc.ProperfoldingenvironmentEfficientsecretionExtra-cellularmatrix–mediumforcellstointeractandmigr

3、ateAnchoragedependantorindependantAnimalcellcultureortheabilitytocontinuouslyculturecellsPlatformtoinvestigatenormalphysiologyandbiochemistryTesttheeffectsofdrugsandcompoundsinvitroTissueengineeringSynthesizevaluableproductsModelsforstudyingdiseasesCellStruc

4、tureWidevarietyofcelllinescultured(epithelial,muscleetc.)Largerthanmicrobialcells(10-30m)ManyinternalorganellesNocellwall–susceptibletoshearforcesCytoskeleton–divisionandmigrationAnchoragedependantCommoncelllinesandtheirapplicationsCellGrowthconditionsGrowt

5、htemperatureMammalian37CInsect27CDissolvedoxygenMediarequirements(insect,mammalian,serumetc.)BasicMediaDifferForMammalianandInsectCultureComponentMammalianInsectGlucose4g/l2.5g/lAminoacids0.01-0.05g/l0.1-0.5g/lGlutamine1g/l1g/lHCO33.5g/l0.35g/lH2PO40.1g/l

6、1g/lSalts4.5g/lNaCl1g/lMg2SO4,KClVitaminsMoreLesspH7.26.4Osmolarity300mOsm350mOsmSerumAdditionUsedat0-20%SerumItcontains:GrowthfactorsTransferrin(Fe)LipidsInsulinAlsoimportantfor:ShearprotectionDetoxificationProblems:Infectiousagents(viruses,mycoplasm,prions

7、)Serumcompositionispoorlydefinedandthebatchesvary.ExpensiveGrowthcurvesinyeastsandmammaliancellsaredifferent0.000010.00010.0010.010.11.010.0050100YeastMammalianMinimumdensityLagphaseExponentialStationaryDeclineCellMetabolismToxicmetabolitesLactatefromglucose

8、metabolism(lowerspH)AmmoniafromcatabolismofglutamateandaminoacidsTypeofcultivationBATCHFERMENTATION.Duration:0.5-1weeksCelldensity4*106c/mlFinish:nomorenutrientsTypeofcultivationFED-BATCHAdditio