Quantitative methods for scoring cell migration and invasion in filter-based assays.
The ability of cells to traverse pores in a biocompatible filter provides means for examining cell chemoattraction. Filter-based assays also permit rapid, quantitative assessment of the in vitro migratory and invasive potential of tumor cells. Scoring migration has relied on visual counting of stained cells which appear on the underside of the filter and determining a true percentage score involves arduous counting of cells on both filter surfaces. Visual counting of random fields may be unreliable, and counting all fields is laborious. In the present study we developed and compared two alternative methods for scoring cell numbers in filter-based assays, a colorimetric assay of toluidine blue binding, and a radioassay of cells prelabeled with [3H]thymidine. Each method was evaluated for sensitivity, variability, ease of use and efficiency, and suitability for use in assays of cell migration and invasion. The radiolabeling method proved to be sensitive and reliable and was the most efficient technique. Although less sensitive and specific, the colorimetric dye method offered a rapid and reliable, nonradioactive alternative with the distinct advantage of preserving intact cultures for follow-up visual assessments. We conclude that colorimetric and radiolabel scoring of filter-based assays are reliable and efficient semiautomated methods which provide means to obtain more complete assessments of cell migration and invasion.
Muir
D
D
Department of Pediatrics, University of Florida College of Medicine, Gainesville 32610.
Sukhu
L
L
Johnson
J
J
Lahorra
M A
MA
Maria
B L
BL
D002452:Cell Count; D002465:Cell Movement; D002633:Chemotaxis; D003124:Colorimetry / Q000379:methods; D005374:Filtration / Q000379:methods*; D005909:Glioblastoma / Q000473:pathology; D006801:Humans; D009361:Neoplasm Invasiveness; D013936:Thymidine; D014048:Tolonium Chloride; D014316:Tritium; D014407:Tumor Cells, Cultured
D014316:Tritium; D014048:Tolonium Chloride; D013936:Thymidine