%0 Journal Article
%T Establishmentandcharacterizationofa?broblast-likecelllinefromAnabariliusgrahami(Cypriniformes:Cyprinidae)
%A Xiaoai WANG
%A Junxing YANG
%A Xiaoyong CHEN
%A Xiaofu PAN
%J 动物学研究
%P 89-97
%D 2012
%R 10.3724/SP.J.1141.2012.E05-06E89
%X ThoughYunnanprovincecontainssome562knownspeciesoffish,nocelllinesfromanyofthesehavebeenmadeavailabletodate.ToprotectgermplasmresourcesandprovideaneffectivetoolinsolvingproblemsatcellularlevelofAnabariliusgrahami,afishendemictoFuxianLake,Yunnan,China,weestablishedandcharacterizedthemajorfeaturesofacontinuouscellline(AGFII)fromthecaudalfintissueofA.grahami.ThisAGFIIcelllineconsistsoffibroblast-likecellsandhasbeensubculturedmorethan60timesoverthecourseofayear.ThecelllinewasmaintainedinDMEM/F12supplementedwith10%FBS,withacellulardoublingtimeof51.1h.Wecontinuedwithmoreexperimentstooptimizethecultureandstorageconditions,andfoundavarietyofinterestingresults:cellscouldgrowattemperaturebetween24癈and28癈,withtheoptimaltemperatureof28癈.Likewise,thegrowthrateofA.grahamifincellsincreasedwhentheFBSproportionincreasedfrom5%to20%,withtheoptimalgrowthattheconcentrationsof20%FBS;cellswereabletogrowinL-15andDMEM/F12withoptimalgrowthatL-15;DMSOisabettercryoprotectantthanGlycerol,EGandMeOHforAGFIIcellswithoptimalconcentrationof5%DMSO.Chromosomeanalysisalsoshowedthatthedistributionofchromosomenumbervariesfrom38to52,withamodalpeakat48chromosomes,accountingfor39.8%ofallcells.UsingthesameprimerpairsspecifictomtDNA,theAGFIIcellsequencesobtainedbyPCRwereidenticaltothosefrommuscletissuesofA.grahami.BothchromosomeanalysisandPCRamplificationconfirmedtheAGFIIcellswerefromA.grahami,alsoindicatingthatthatcurrentlong-termartificialpropagationofA.grahamihasbeensuccessful.Finally,wenotedthatwhencellsweretransfectedwithpEYFP-N1andpECFP-N1plasmid,brightfluorescentsignalswereobserved,suggestingthatthiscelllinemaybesuitableforuseintransfectionandfuturegeneexpressionstudies.
%K Characterization
%K Fibroblast-likecellline
%K Anabariliusgrahami
%K Cryopreservation
%U http://www.zoores.ac.cn/CN/abstract/abstract3263.shtml