Materialstobepreparedbeforehand:1)FBSfreemedium2)10%FBSmedium3)Cellmigrationfilterinsert(Transwell®,12mmDiameter,12μmPoreSize.)Procedures:1)Coattheundersideoffilterinsertswith20μg/mloffibronectinovernightat4oCAirdrytheinsertbeforethecellswereadded.2)TrypsinizecellsandwashtwicewithFBSfreemedium.3)Countcellsto4×105/ml.4)Add0.5mlcellsinFBSfreemedium10thetopofinsert.5)Add1mlofmedium(with10%FBSand10μg/mlCollagenI)tothelowerchamber.6)Putfilterinsertintothelowerchamberandincubateovernightat37oC.7)RemovecellsonthetopsideofthefilterbyscrubbingtwicewithcottontippedswabmoistenedwithnoFBSmedium.8)StaincellswithDiff-QuikTMstainkit.9)Washwithdistilledwater.10)Mount.