Haploid plants achieve through androgenesis or gynogenesis. In gynogenesis method, the ovary or ovule are used as explants induct haploid plants. Female flower one day before flowering of Cucumis sativus L. are collected. Cold pretreatment of ovaries at 4°C up to 24 hours and culture under dark conditions. Significantly enhanced callus induction response is compared with cultures under 4-week cultured on CBM medium supplemented with various concentration of TDZ 0.01-0.04 mg/L. After 4 weeks, ovaries are transferred to medium with kinetin 0.05 – 0.20 mg/L. Then, ovaries were transferred to medium supplemented with BA: IAA 3:1. Finally, green ovaries were transferred to BA 1.5 mg/L and GA3 1.5 mg/L. The results showed that ovary induction has best affected on CBM with TDZ 0.03 mg/L with 11 callus/sample. Ovaries developed on kinetin 0.1 mg/L with 7.4 callus/sample. Ovaries become green and had leaves and roots formation on BA: IAA (3 mg/L: 1 mg/L). 11 plantlets were harvested from ovary culture after 12-week culture on CBM supplemented with BA 1.5 mg/L and GA3 1.5 mg/L.