Artificial cell-spreading of permeabilized BHK cells on fibronectin-coated substratum

We developed a cell-spreading model of permeabilized BHK cells to analyze molecular interactions in the cell-spreading machinery. BHK cells in suspension were allowed to attach onto the fibronectin-coated cover slips at 37 degrees C for 20 min. The cells were then permeabilized with 50 micrograms/ml saponin for 10 min at 4 degrees C. Approximately 50 percent of the cell models spread in response to exogenous 1 mM ATP for 90 min at 37 degrees C. Anti-actin antibody penetrated into almost all the cell models which spread ATP-dependently. These indicate that half number of the cell models permeabilized with saponin is spread by the exogenous ATP and has pores larger than an IgG molecule.