Evidence for two distinct mechanisms of anchorage stimulation in freshly explanted and 3T3 Swiss mouse fibroblasts

When single cells were allowed to attach to circular islands of adhesive substratum, their proliferation was strongly dependent on island area over the range 500 micron2 to 5000 micron2. The number of freshly explanted whole mouse embryo fibroblasts that performed DNA synthesis corresponded closely with a simple geometrical measure of area of cell surface exposed to the medium: freely suspended cells were only slightly less stimulated than attached cells exposing an equal surface area; hemispherical cells were less stimulated than cells of any other shape. In contrast, 3T3 cells were stimulated sixfold by islands too small to allow any increase in area. These experiments show that anchorage can stimulate by two different mechanisms. They offer a general method of measuring substrate contact stimulation.

[1]  H. Green,et al.  QUANTITATIVE STUDIES OF THE GROWTH OF MOUSE EMBRYO CELLS IN CULTURE AND THEIR DEVELOPMENT INTO ESTABLISHED LINES , 1963, The Journal of cell biology.

[2]  K. Crossin,et al.  Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis , 1981, Cell.

[3]  M. Stoker,et al.  Shaking 3T3 cells: further studies on diffusion boundary effects. , 1974, Cell.

[4]  M. Moskowitz,et al.  Arrest of 3T3 cells in G1 phase in suspension culture , 1976, Journal of cellular physiology.

[5]  L. Stolt,et al.  Proliferation control in cloned normal and malignant human cells. , 1980, Experimental cell research.

[6]  G. Dunn,et al.  New evidence that growth in 3T3 cell cultures is a diffusion-limited process , 1984, Nature.

[7]  B. Westermark,et al.  Growth control in miniclones of human glial cells. , 1978, Experimental cell research.

[8]  A. Ben-Ze'ev,et al.  Multinucleation and inhibition of cytokinesis in suspended cells: Reversal upon reattachment to a substrate , 1981, Cell.

[9]  J. Campisi,et al.  Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum , 1983, Journal of cellular physiology.

[10]  J. Folkman,et al.  Role of cell shape in growth control , 1978, Nature.

[11]  E. Rozengurt,et al.  Stimulating the proliferation of quiescent 3T3 fibroblasts by peptide growth factors or by agents which elevate cellular cyclic AMP level has opposite effects on motility. , 1985, Experimental cell research.

[12]  B G Nordestgaard,et al.  Critical‐point drying versus freeze drying for scanning electron microscopy: a quantitative and qualitative study on isolated hepatocytes , 1985, Journal of microscopy.

[13]  E. Rozengurt,et al.  The role of cytoplasmic microtubules in the regulation of the activity of peptide growth factors. , 1980, Advances in enzyme regulation.

[14]  R. Brooks,et al.  Apparent heterogeneity in the response of quiescent swiss 3T3 cells to serum growth factors: Implications for the transition probability model and parallels with “cellular senescence” and “competence” , 1984, Journal of cellular physiology.

[15]  C. O'neill,et al.  Anchorage and growth regulation in normal and virus‐transformed cells , 1968, International journal of cancer.

[16]  R. Shields,et al.  Cell growth, cell division and cell size homeostasis in Swiss 3T3 cells. , 1985, Experimental cell research.

[17]  N. G. Maroudas Appendix: Short-range diffusion gradients , 1974 .

[18]  B. Tighe,et al.  Requirements for cell spreading on polyHEMA coated culture substrates. , 1984, Cell biology international reports.

[19]  J. Trinkaus,et al.  Microvilli and blebs as sources of reserve surface membrane during cell spreading. , 1976, Experimental cell research.

[20]  M. G. P. STOKER,et al.  Role of Diffusion Boundary Layer in Contact Inhibition of Growth , 1973, Nature.

[21]  R. Goldman,et al.  The occurrence of microvilli during spreading and growth of BHK21-C13 fibroblasts. , 1970, Experimental cell research.

[22]  A. Boyde,et al.  Freeze‐drying shrinkage of glutaraldehyde fixed liver , 1981, Journal of microscopy.

[23]  Sheldon Penman,et al.  Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts , 1980, Cell.

[24]  A. W. Rogers Techniques of autoradiography , 1967 .

[25]  T. Hunter,et al.  Vinculin: A cytoskeletal target of the transforming protein of rous sarcoma virus , 1981, Cell.

[26]  A. Ben-Ze'ev,et al.  The cytoskeleton in cancer cells. , 1985, Biochimica et biophysica acta.

[27]  J. Folkman,et al.  Influence of geometry on control of cell growth. , 1975, Biochimica et biophysica acta.

[28]  P. Riddle,et al.  The relation between surface area and anchorage dependence of growth in hamster and mouse fibroblasts , 1979, Cell.

[29]  J. Smith,et al.  Growth factors adherent to cell substrate are mitogenically active in situ , 1982, Nature.