资源描述：
《Changing medium and passaging cell lines》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。
1、PROTOCOLChangingmediumandpassagingcelllinesJohnRMasters1&GlynNStacey21DepartmentofSurgery,ProstateCancerResearchCentre,RoyalFreeandUniversityCollegeLondonMedicalSchool,67RidingHouseStreet,LondonW1W7EJ,UK.2NationalInstituteforBiologicalStandardsandControl,BlancheLane,SouthMimms,Potter
2、sBar,HertfordshireEN63QG,UK.CorrespondenceshouldbeaddressedtoJ.R.M.(j.masters@ucl.ac.uk).Publishedonline13September2007;doi:10.1038/nprot.2007.319Celllinesarewidelyusedinbiomedicalresearch.Thisprotocoldescribesthemethodsusedroutinelytochangethemediumandpassagethecells.Mediumchangeske
3、epthecellshealthybyprovidingfreshnutrients,whilecellpassageorsplittingisrequiredtosmaintaincellsinexponentialgrowth.Despitethesimplicityofthemethodsused,eachcelllinehasidiosyncracies.Whetherworkingwithoneorseveralcelllines,thereisnosubstituteforknowledgeoftheirneeds,includingtherange
4、ofphenotypesandgrowthpatternsunderdifferentphysicalandnutrientconditions.Giventhenecessarycareandattention,mostcelllinesareeasytomaintainandgrow.natureprotocol/mINTRODUCTIONoc.Fiftyyearsago,maintenanceofcelllineswasarduous.Tissuelineismaintainedinculture,thegreaterthechancethatitwill
5、erculturemediumhadtobemadefrombaseconstituents,serumacquireoneormorechangesingenotype.Suchchangesmayutawascollectedfromtheabattoir,therewerenotissuecultureplasticscriticallyalterthecharacteristicsofthecellline,butwecann.wandsterilitywasmaintainedontheopenbenchwithoutlaminarminimizege
6、neticchangeandothervariablesinfluencingthecellsww/flowhoodsorantibiotics.However,asdescribedinthisprotocol,byobservingstrictadherencetogoodcellculturepractice.The/:ptmanyoftheissuesthatresearchersfaced50yearsagostillneedtobemostimportantmeansofminimizingsuchchangesistorestrictthethcons
7、ideredtodaytoobtainreproducibleandmeaningfuldata.lengthoftimethatcelllinesaregrownincontinuousculture.AsapEachandeverycelllinehasslightlydifferentrequirements,pragmaticruleofthumb,weadvocatethatacelllineshouldneveruorphenotypeandbehavior,andthereisnosubstituteforfamiliaritybeincontin
8、uousculturef
