Effect of Substratum Morphology on Animal Cell Adhesion and Behavior
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EFFECT OF SUBSTRATUM MORPHOLOGY ON ANIMAL CELL ADHESION AND BEHAVIOR RAHUL SINGHVI, GREGORY N. STEPHANOPOULOS AND DANIEL I. C. WANG Biotechnology Process Engineering Center and Department of Chemical Engineering Massachusetts Institute of Technology, 77 Massachusetts Avenue Cambridge, Massachusetts 02139. ABSTRACT Glass surfaces with well defined surface morphologies have been prepared using photolithography to study the effect of surface morphology on cell adhesion and function. Using a transformed recombinant cell-line, AtT-20, as a model of shear sensitive cell, we have shown that cell-substratum adhesion strength is enhanced using a surface with uniform grooves without any loss in cellular function. Furthermore, using primary hepatocytes as a model for a cell whose function is sensitive to its shape, we have shown that surface morphology can modulate cell shape as well as its function. INTRODUCTION Large-scale cultivation of animal cells is becoming increasingly important for the production of complex proteins such as erythropoietin, tissue plasminogen activator and monoclonal antibodies. As manufacturing of these proteins becomes more competitive, highly efficient culturing systems need to be developed. Furthermore, in any such system, effects of the cell's microenvironment will have to be elucidated which could lead to a better understanding on the microheterogeneities in the protein product. It is with this theme in mind we have investigated the use of porous ceramic particles for large-scale animal cell culture. In the course of this study, we have observed that the ceramic surface is rough and that cell shape and spreading seem to be affected by differences in the substratum surface morphology. Several recent studies have demonstrated that cell shape (of normal cells) can affect cell growth and function [1,2,3,4]. Differences in cell shape can alter the cytosketon architecture which, in turn, can affect gene expression [5]. In light of these findings, we have hypothesized that surface morphology could affect cell shape and spreading and possibly cell adhesion and function. To test our hypotheses, we have directed our efforts to systematically elucidate the effects of substratum morphology on animal cell adhesion and behavior. High quality glass surfaces with well defined surface morphologies have been used in aid of our research. Glass was chosen as the substrate since it is a well accepted cell culture substrate and is easy to modify to produce well defined surface morphologies using photolithography and other techniques developed in microelectronics processing. Furthermore, the transparent property of glass allows cells to be easily visualized and monitored at various stages of culture. Effects of surface morpholgy on cell surface adhesion strength, cell shape and spreading and protein secretion characteristics have been measured using suitable assays with smooth glass surfaces as control. Here, we report some of our initial findings.
Mat. Res. Soc. Symp. Proc. Vol. 252. 'c1992 Materials Research Societ
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