Materials
FibroblastsUSPensioncultureTissueculturelaminarflowhoodMediaappropriatetoculturelineusedDisposablePipettes(10mland1.0ml)DisposablecultureflasksProcedure
Obtainacultureofmousefibroblastcellsinsuspensionculture.Thiswillbeasimpleculturewithminimalrequirements,andoneselectedforexcellentgrowthcharacteristics.Thetransferprocedurewillbesimilartothatforprokaryotes,withafewmajorchanges.First,alltransferswillbedoneinatissueculturehoodinordertomaximizeasepsis.Secondly,thecellswillbetransferredwithsiliconizedpipettesratherthanwireloops.Thesiliconepreventsadheranceofthecellstotheglasswallofthepipette.Atissueculturehoodisadevicethathasairmovinginlayersandunderpositivepressure.Sincetheairisfiltered,itcontainsminimalnumbersofbacteriaorfungalspores,andsinceitisunderapositivepressure,thoseparticlesthatarepresentareblownoutofthehood.Thelayeringpreventsairborneorganismsfromsettlingontheworksurfaces.
Arrangethematerialsinfrontofyou,easilyaccessIBLethroughtheopeningofthetissueculturehood.Ensurethatanyalcoholsandwrappingpaperarekeptclearofthebunsenburner.Pre-sterilizethehoodbeforeuse,andusedisposablesterilegloves.Loosenthecapofatissuecultureflaskandthecapofastockbottleoftissueculturemedia.6Insertthetipofasterilepipetteintothestockbottleandremove10mlofmedia.Transferthemediatothetissuecultureflask.
Openthetopofthesuspensioncultureanduseasterile1.0mltransferpipettetoremovea1.0mlsampleoftheculture.Transferittothefreshmediainthecultureflask.Secureallcapsthathavebeenloosened.Placethenewculturesinanincubatorat37°C.
>>> 更多资讯详情请访问蚂蚁淘商城
