Characterization of Growth Properties through Morphological Observation in Culture of Anchorage-Dependent Cells

Abstract

The focus on the end point of the sequence of intracellular reactions, namely, the behavioral phenotypes has been dedicated to emerging technologies for characterizing motility-related parameters. In the present study, the cell attachment and spreading on culture surface during the adhesion phase after inoculation were examined and a morphological rate parameter was proposed for the evaluation of growth potential during the culture of mouse fibroblast cells (3T3) using the tool developed for the observation of cell morphology. To ensure the validity of the tool, the adherent area of an individual cell was measured during the culture. After inoculation, it was observed that cells attached and then spread on the bottom surface of T-flask. The adherent areas of cells on the surface, which were determined using our system, expanded with elapsed time, approaching to a saturated value of the area. After that, cell division occurred with a sharp decrease in the area. In addition, re-spreading of the individual cell was observed, indicating that our system was available for measurement of the cell spreading and cell division. By using the system, the variation in adherent area of an individual cell was investigated with the cells that underwent after the treatment for cell detachment with trypsin solution for given periods. The excessive treatment resulted in a decrease in spreading rate of cells. Moreover, the time of first cell division decreased with increasing spreading rate, suggesting that the spreading rate was an available parameter to evaluate the growth potential.