Correlation of the cell phenotype of cultured cell lines with their adhesion to components of the extracellular matrix☆

摘要

Thedifferentialadhesionofculturedmammalianclonalcelllinestocomponentsoftheextracellularmatrixwasexaminedbykineticadhesionandlong-termgrowthassays.UniformartificialmatriceswerepreparedbyairdryingcollagenTypeIsolution(C)ontoamicrotiterwellandthenairdryingasolutioncontainingasingleglycosaminoglycan(GAG):hyaluronicacid(HA),chondroitinsulfate-4(CHS-4),orchondroitinsulfate-6(CHS-6).Theadhesionof[3H]thymidine-prelabeledcellssuspendedinfibronectin(FN)depletedmediumwasmeasuredat2and6hr.Neuroblastoma(N18,Lan1)andmelanoma(B16,G361,S91)celllinesexhibitedasignificantlygreaterpercentageofcellsadheringtooneormoreC-GAGmatricescomparedwithCmatrices.Maximaladhesionat2hrwastoC-HA.Incontrastat2hr,twoglial,twoepithelial,andonefibroblasticcelllineshowedunchangedorsignificantlydecreasedbindingtoC-GAGcomparedwithCmatrices.Furtherexperimentsusinganeuroblastoma(N18)andaglioma(C6)celllineindicatedthattheadhesionpatternswerenotalteredeitherbythemethodofdissociationfromthetissueculturedish,preincubationwithexogenousGAG,ortheadditionofexogenousfibronectin.AssaysofN18andC6adhesiontomatricesmadefromanon-GAGpolyanioniccompound,polygalacturonicacid(PGA),didnotyieldthesameadhesionpatternsasC-HAmatrices.Long-termgrowthstudiesofaneuroblastoma(N18)melanoma(S91),andglioma(C6)celllineonnonuniformmatricesdeliberatelypreparedwithGAG-richandGAG-poorregionscomplementedtheobservationsfromthekineticadhesionassays.N18andS91cellsdidnotgrowonareaswhichdidnotcontainGAGbytoluidinebluestaining.However,theC6cellsdidnotgrowonareaswhichdidstronglystainforGAG.AquantitativeanalysisofthelongtermgrowthofN18andC6cellssubstantiatedtheseobservations.Allthesedataindicatethatthecellularphenotypemaybecorrelatedwithmatrixadhesion.NeuroblastomasandmelanomashaveagreateraffinityforGAG-containingmatriceswhileglial,epithelial,andfibroblasticcellsappeartohaveagreaterorequalaffinityforcollagenmatrices.