ForResearchUseOnly.Notforuseindiagnosticortherapeuticprocedures.BDBiosciencesAssayMethodsProtocol:CellInvasionAssayTheInvasionAssayprovidesaninvitrosystemtostudycellinvasionofmalignantandnormalcells.Specificapplicationsincludeassessmentofthemetastaticpotentialoftumorcells1,inhibitionofmetastasisbyextracellularmatrixcomponents2orantineoplasticdrugs(TAXOL®)3,alteredexpressionofcellsurfaceproteins4,ormatrixmetalloproteinases5inmetastaticcells;andinvasionofnormalcellssuchasembryonicstemcells6,cytotrophoblasts7,endothelialcells8,andfibroblasts9.Invasionstudieshavebeensuccessfullyperformedonavarietyoftumorcells(celllinesandprimarytumors)includingmelanomas,glioblastomas,astrocytomas,osteosarcomas,fibrosarcomas,andadenocarcinomasofthelung,prostate,breast,ovary,andkidney.InvasionchamberscoatedwithBDMatrigel™matrixprovidecellswiththeconditionsthatallowassessmentoftheirinvasivecapacityinvitro.BDMatrigelmatrixservesasareconstitutedbasementmembraneinvitro,occludingtheporesofthemembraneandblockingnon-invasivecellsfrommigratingthroughthemembrane.Incontrast,invasivecells(malignantandnon-malignant)secreteproteasesthatenzymaticallydegradetheBDMatrigelmatrixandenableinvasionthroughthemembranepores.Themembranemaybeprocessedforlightandelectronmicroscopyandcanbeeasilyremovedafterstaining.ThefollowingprocedurehasbeenoptimizedusingHT-1080humanfibrosarcomacellsonBDMatrigelmatrix-coatedinserts(8µmporediameter).Resultsmayvarydependingupontheexperimentalconditionsincludingthecells10,medium,incubationtime,cellseedingdensity,andchemoattractant.Therefore,theconditionsincludingcellcultureinsertporesize10shouldbeoptimizedforeachsystem.Materials:•BDBioCoat™cellcultureinsertspackagedincompanionplates(24-well,BDcat.no.354578),orindividualinserts(24-well,BDcat.no.353097).BDFalcon™cellcultureinsertsarealsoavailableinvariousporesizes.•BDFalcon™cellcultureinsertcompanionplates(24-well,BDcat.no.353504).•BDMatrigelbasementmembranematrix(10mlvial;standardBDMatrigel,BDcat.no.354234;orGrowthFactorR...
