Abstract: There were a lot of free vacuoles to have been examinated in the liquidcultrues which had been preserved for over 3 months. The vacuoles were typicalyspherical, fully transparent and varied in size. The giant vacuoles showed aphagocytosis and there were many aged cells to be engulfed by them. Both the agedand nonaged cells which were grown for 3 months, 2 months, 1 month, 18 days, 10days and 3 days could also be able to release vacuoles in the isolation experiments.Three methods, enzymolysis in hypotonic condition, osmotic shock and hypotonicenzymolysis-osmotic shock, were used for the isolation of vacuoles. Over 15% ofcells which were grown for 3 days could release vacuoles in the experiments.