The effect of substrate and adsorbed proteins on adhesion, growth and shape of CaCo‐2 cells

Abstract

Extracellular matrix (ECM) proteins play a critical role in many cellular functions, from spreading, migration and proliferation to apoptosis. This role can be altered when proteins of the native ECM are adsorbed to different substrates which cause structural modifications that can influence their biological function. The effects on CaCo-2 cells of laminin-1, fibronectin, collagen-1 and ECM gel adsorbed to glass and to tissue culture polystyrene (PS) were compared in terms of adhesion, proliferation, shapes and spreading of cells in culture. Significant differences between glass and PS surfaces were observed for proliferation and cell shape. Protein surfaces prepared on PS substrates had, in most cases, more pronounced effects on cells than uncoated PS, especially if coated by collagen-1. Adsorbed ECM gel was the most adhesive for cells, but its effect on cell proliferation was not notably different from the controls (glass or PS). These findings indicate that the choice of the substrate can have a significant effect on experimental results and should be taken into consideration when comparing results obtained on different surfaces.

Time Lapse Monitoring of CaCo-2 Cell Shapes and Shape Dependence of the Distribution of Integrin β1 and F-Actin on their Basal Membrane

“…Noncoated substrates displayed the slowest rate of shape changes. All these results are in accordance with the results on fixed cells (Zemljič Jokhadar et al 2007); however, the time lapse measurements provide additional information on intermediate shape changes in terms of the occurrence of type 2* cells and the transitional role of type 2 cells.…”

“…Therefore, in order to get direct information on the succession of cell shape changes, we decided to monitor live cells in this study. The imaging of live cells is a noninvasive method but gave a less detailed view of cells than was obtained with fixed cells, especially in the case of spread cells, and types 2 and 3 of the previous classification (Zemljič Jokhadar et al 2007) could not be distinguished. Both these types were therefore joined here into a single cell type 2 (Figure 1).…”

“…CaCo-2 epithelial cell line was used as a model to predict epithelial cell behavior in general. In a previous study (Zemljič Jokhadar et al 2007), we succeeded to link CaCo-2 cell shapes with cell processes such as adhesion and proliferation, using cells fixed at various times after seeding. The imaged cells were categorized into five shape types, the proportion of which was obtained at each time point.…”

“…The values for all parameters studied were significantly higher on PS surfaces than those for glass surfaces. On collagen-1 coatings, cells adhered, spread, and reached the end shape state faster than on laminin-1, fibronectin, or noncoated substrates (Zemljič Jokhadar et al 2007). …”

“…For characterization of the stability of CaCo-2 cell populations (either of parental or clonal origin) in terms of morphology, growth characteristics, and differentiation, it is important to monitor the fate of individual cells. We recently introduced a method (Zemljič Jokhadar et al 2007) by which we followed the fate of cells that grow on different substrates as early as possible after being seeded. The purpose was to link the shapes of CaCo-2 cells observed over a short time post seeding and processes such as adhesion and proliferation later in the life of cells.…”