The constant-current chronopotentiometric measurements of egg yolk phosphatidylcholine bilayer membrane without and with cholesterol are presented. It was demonstrated that the constant-intensity current flow through the bilayer membranes without and with cholesterol generated the oscillating pores in their structures. The presence of cholesterol in the bilayer membrane increased the value of critical potential at which pores could be formed. The shift in the distribution of calculated pore radii towards smaller values was observed in the bilayer containing cholesterol. It was postulated that greater stability of bilayer with cholesterol resulted from increased critical pore radius (at which the bilayer would rupture). The implications of the membrane cholesterol for the application of constant-current method as a biotechnological tool for incorporating molecules of different size are discussed.