Characterization of Cellular Motions through Direct Observation of Individual Cells at Early Stage in Anchorage-Dependent Culture

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In the monolayer culture of murine fibroblasts, the characterization of cellular motions was performed by morphological observation using a tool consisting of an optical assembly connected to a computer-aided image analysis system. This tool enabled the estimation of the projected area of the cells with sufficient accuracy, being able to follow the serial behavior of the cells on the culture surface. Trypsinization was chosen as an external factor for altering the state of the cells, and the rate of cell spreading (r_s) and time of first cell division (t_1) were evaluated by subjecting the cells to trypsin treatment for t_T=3 and 15 mm. During culture of the cells treated for t_T=3 mm, the r_s values of individual cells exhibited a broad variation, ranging from 41 to 321 ★m^2/h, and the average r_s was 165★78 ★m^2/h, which was 1.5 times larger than that of the cells treated for t_T=15 mm. On the contrary, the average t_1 of the cells treated for t_T=3 mm was 9.5 h which was 60% reduced as compared with that of the cells treated for t_T=15 mm. The prolonged time of trypsin treatment was considered to induce the decrease in r_s and delay of the first cell division due to the requirement for the recovery from cell surface damage caused by trypsinization. The logarithmic plots of r_s and t_1, were found to have an inverse relation regardless of the state of the individual cells.
社団法人日本生物工学会の論文
2002-10-25