JournalofVisualizedExperimentswww.jove.comCopyright©2011JournalofVisualizedExperimentsandCorning,Inc.December2011|58|e3418|Page1of6VideoArticleScale-UpofMammalianCellCultureusingaNewMultilayeredFlaskElizabethJ.Abraham1,KatieA.Slater1,SuparnaSanyal1,KenLinehan1,PaulaM.Flaherty1,SusanQian11BDBiosciencesURL:http://www.jove.com/video/3418DOI:doi:10.3791/3418Keywords:BasicProtocols,Issue58,Multi-Flask,multi-layered,stackable,scale-up,cellculture,flasksDatePublished:12/5/2011Citation:Abraham,E.J.,Slater,K.A.,Sanyal,S.,Linehan,K.,Flaherty,P.M.,Qian,S.Scale-UpofMammalianCellCultureusingaNewMultilayeredFlask.J.Vis.Exp.(58),e3418,doi:10.3791/3418(2011).AbstractAgrowingnumberofcell-basedapplicationsrequirelargenumbersofcells.UsageofsinglelayerT-flasks,thatareadequateduringsmall-scaleexpansion,maybecomecumbersome,laboriousandtime-consumingwhenlargenumbersofcellsarerequired.Toaddressthisneed,theperformanceofanewmulti-layeredcellculturevesseltofacilitateeasyscaleupofcellsfromsinglelayeredT-flaskswillbediscussed.Theflaskstestedareavailablein3-and5-layerformatandenablecultureandcompleterecoveryofthreeandfivetimesthenumberofcellsrespectively,comparedtoT-175flasks.AkeyfeatureoftheBDMulti-Flaskisamix/equilibrationportthatallowsrapidin-vesselmixingaswellasuniformdistributionofcellsandreagentswithinandbetweenlayersofeachvesselandconsistentlyproducecellsthatcanbeculturedinanenvironmentthatiscongruenttoT-175flasks.ThedesignoftheseMulti-Flasksalsoallowsforconvenientpipetteaccessforaddingreagentsandcellsdirectlyintotheflasksaswellasefficientrecoveryofvaluablecellsandreagentsandreducesriskofcontaminationduetopouring.Forapplicationswherepouringispreferredoverpipetting,thedesignallowsforminimalresidualliquidretentionsoastoreducewastageofvaluablecellsandreagents.VideoLinkThevideocomponentofthisarticlecanbefoundathttp://www.jove.com/video/3418/Protocol1.ProtocoltouseMulti-Flasksforcellculture1.Preparationofvesselsandaddingcellsuspension1.Preparecellgrowthmediumasneeded.2.LineupMulti-Flaskver...
