细胞培养方法

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1、12.0   CellCultureProtocols12.1   BasicTechniques-The"Do'sandDon'ts"ofCellCultureGivenbelowareafewoftheessential"do’sanddon'ts"ofcellculture.Someofthesearemandatorye.g.useofpersonalprotectiveequipment(PPE).Manyofthemarecommonsenseandapplytoalllaboratoryareas.Howeversomeofthemarespecifictoti

2、ssueculture.TheDo’s1.Usepersonalprotectiveequipment,(laboratorycoat/gown,glovesandeyeprotection)atalltimes.Inaddition,thermallyinsulatedgloves,full-facevisorandsplash-proofapronshouldbewornwhenhandlingliquidnitrogen.2.Alwaysusedisposablecapstocoverhair.3.WeardedicatedPPEfortissueculturefaci

3、lityandkeepseparatefromPPEworninthegenerallaboratoryenvironment.Theuseofdifferentcoloredgownsorlaboratorycoatsmakesthiseasiertoenforce.4.Keepallworksurfacesfreeofclutter.5.Correctlylabelreagentsincludingflasks,mediumandampuleswithcontentsanddateofpreparation.6.Onlyhandleonecelllineatatime.T

4、hiscommon-sensepointwillreducethepossibilityofcrosscontaminationbymislabelingetc.Itwillalsoreducethespreadofbacteriaandmycoplasmabythegenerationofaerosolsacrossnumerousopenedmediabottlesandflasksinthecabinet.7.Cleantheworksurfaceswithasuitabledisinfectant(e.g.70%ethanol)betweenoperationsand

5、allowaminimumof15minutesbetweenhandlingdifferentcelllines.8.Whereverpossiblemaintainseparatebottlesofmediaforeachcelllineincultivation.9.Examineculturesandmediadailyforevidenceofgrossbacterialorfungalcontamination.Thisincludesmediumthathasbeenpurchasedcommercially.10.QualityControlallmediaa

6、ndreagentspriortouse.11.Keepcardboardpackagingtoaminimuminallcellcultureareas.12.Ensurethatincubators,cabinet,centrifugesandmicroscopesarecleanedandservicedatregularintervals.13.Testcellsformycoplasmaonaregularbasis.TheDon’ts1.Donotcontinuouslyuseantibioticsinculturemediumasthiswillinevitab

7、lyleadtotheappearanceofantibioticresistantstrainsandmayrenderacelllineuselessforcommercialpurposes.2.Don’tallowwastetoaccumulateparticularlywithinthemicrobiologicalsafetycabinetorintheincubators.3.Don'thavetoomanypeopleinthelabatanyonetime.1.Don'thandlec