TY - JOUR

T1 - Individual cell motility studied by time-lapse video recording: influence of experimental conditions.

AU - Hartmann-Petersen, R

AU - Walmod, P S

AU - Berezin, A

AU - Berezin, V

AU - Bock, E

N1 - Keywords: Actins; Animals; Cell Count; Cell Movement; Cells, Cultured; Fibroblasts; Fibronectins; Hydrogen-Ion Concentration; L Cells (Cell Line); Mice; Microscopy, Confocal; Microscopy, Phase-Contrast; Osmolar Concentration; Temperature; Video Recording

PY - 2000

Y1 - 2000

N2 - BACKGROUND: Eukaryotic cell motility plays a key role during development, wound healing, and tumour invasion. Computer-assisted image analysis now makes it a realistic task to quantify individual cell motility of a large number of cells. However, the influence of culture conditions before and during measurements has not been investigated systematically. METHODS: We have evaluated intraassay and interassay variations in determinations of cellular speed of fibroblastoid L929 cells and investigated the effects of a series of physical and biological parameters on the motile behavior of this cell line. Cellular morphology and organization of filamentous actin were assessed by means of phase-contrast and confocal laser scanning microscopy and compared to the corresponding motility data. RESULTS: Cell dissociation procedure, seeding density, time of cultivation, and substrate concentration were shown to affect cellular speed significantly. pH and temperature of the medium most profoundly influenced cell motility and morphology. Thus, the mean cell speed was 40% lower at pH 7.25 than at pH 7.6; at 29 degrees C, it was approximately four times lower than at 39 degrees C. CONCLUSION: Of the parameters evaluated, cell motility was most strongly affected by changes in pH and temperature. In general, changes in cell speed were accompanied by alterations in cell morphology and organization of filamentous actin, although no consistent phenotypic characteristics could be demonstrated for cells exhibiting high cell speed.

AB - BACKGROUND: Eukaryotic cell motility plays a key role during development, wound healing, and tumour invasion. Computer-assisted image analysis now makes it a realistic task to quantify individual cell motility of a large number of cells. However, the influence of culture conditions before and during measurements has not been investigated systematically. METHODS: We have evaluated intraassay and interassay variations in determinations of cellular speed of fibroblastoid L929 cells and investigated the effects of a series of physical and biological parameters on the motile behavior of this cell line. Cellular morphology and organization of filamentous actin were assessed by means of phase-contrast and confocal laser scanning microscopy and compared to the corresponding motility data. RESULTS: Cell dissociation procedure, seeding density, time of cultivation, and substrate concentration were shown to affect cellular speed significantly. pH and temperature of the medium most profoundly influenced cell motility and morphology. Thus, the mean cell speed was 40% lower at pH 7.25 than at pH 7.6; at 29 degrees C, it was approximately four times lower than at 39 degrees C. CONCLUSION: Of the parameters evaluated, cell motility was most strongly affected by changes in pH and temperature. In general, changes in cell speed were accompanied by alterations in cell morphology and organization of filamentous actin, although no consistent phenotypic characteristics could be demonstrated for cells exhibiting high cell speed.

M3 - Journal article

C2 - 10918277

VL - 40

SP - 260

EP - 270

JO - Cytometry Part B - Clinical Cytometry

JF - Cytometry Part B - Clinical Cytometry

SN - 1552-4949

IS - 4

ER -