Abstract:Using hypocotyls (5～10 mm) of Apium graveolens L. as explant, calli were induced in induction medium (MS + 1.0 mg/L 2, 4-D). The embryogenic calli were transformed to differentiation medium (MS+0. 5 mg/L kinetin+ 500 mg/L CH+500 mg/L Prolin) after several subsequent subcultures and selection by replacement of solid and liquid medium. Technical conditions such as the shake rate of the flask, the initial cell density, as well as subsequent the initial pH values during culture were under consideration. With the optimum flask shake rate of about 100～150 r/min, initial cell density of 2.0% (fresh weight) and the initial pH value of 5.5, the authors have obtained 130 normal cotyledon embryos in each mL of cultures.