Changing medium and passaging cell lines(2)

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1、PROTOCOLChangingmediumandpassagingcelllinesJohnRMasters1&GlynNStacey21DepartmentofSurgery,ProstateCancerResearchCentre,RoyalFreeandUniversityCollegeLondonMedicalSchool,67RidingHouseStreet,LondonW1W7EJ,UK.2NationalInstituteforBiologicalStandardsandControl,BlancheLane,SouthMimms

2、,PottersBar,HertfordshireEN63QG,UK.CorrespondenceshouldbeaddressedtoJ.R.M.(j.masters@ucl.ac.uk).Publishedonline13September2007;doi:10.1038/nprot.2007.319Celllinesarewidelyusedinbiomedicalresearch.Thisprotocoldescribesthemethodsusedroutinelytochangethemediumandpassagethecells.M

3、ediumchangeskeepthecellshealthybyprovidingfreshnutrients,whilecellpassageorsplittingisrequiredtosmaintaincellsinexponentialgrowth.Despitethesimplicityofthemethodsused,eachcelllinehasidiosyncracies.Whetherworkingwithoneorseveralcelllines,thereisnosubstituteforknowledgeoftheirne

4、eds,includingtherangeofphenotypesandgrowthpatternsunderdifferentphysicalandnutrientconditions.Giventhenecessarycareandattention,mostcelllinesareeasytomaintainandgrow.natureprotocol/mINTRODUCTIONoc.Fiftyyearsago,maintenanceofcelllineswasarduous.Tissuelineismaintainedinculture,t

5、hegreaterthechancethatitwillerculturemediumhadtobemadefrombaseconstituents,serumacquireoneormorechangesingenotype.Suchchangesmayutawascollectedfromtheabattoir,therewerenotissuecultureplasticscriticallyalterthecharacteristicsofthecellline,butwecann.wandsterilitywasmaintainedont

6、heopenbenchwithoutlaminarminimizegeneticchangeandothervariablesinfluencingthecellsww/flowhoodsorantibiotics.However,asdescribedinthisprotocol,byobservingstrictadherencetogoodcellculturepractice.The/:ptmanyoftheissuesthatresearchersfaced50yearsagostillneedtobemostimportantmeansof

7、minimizingsuchchangesistorestrictthethconsideredtodaytoobtainreproducibleandmeaningfuldata.lengthoftimethatcelllinesaregrownincontinuousculture.AsapEachandeverycelllinehasslightlydifferentrequirements,pragmaticruleofthumb,weadvocatethatacelllineshouldneveruorphenotypeandbehavi

8、or,andthereisnosubstituteforfamiliaritybeincontinuousculturef