This study was performed to develop the mass propagation system using tissue culture technique to supply the seeds of Elephant garlic (Allium ampeloprasum L.) which has difficulty in propagation. Immature spathe of Elephant garlic was cultured on Murashige & Skoog (MS) medium supplemented with two plant growth regulators, naphthaleneacetic acid (NAA) and kinetin. After 6 weeks of culture, the highest number of shoot (14.9/explant) was obtained when the immature spathe with 10 ㎝ length was cultured right after harvesting. In MS medium supplemented with 2 ㎎/L kinetin and 0.5 ㎎/L NAA, the most vigorous growth characteristics was observed, the shoot number was 14.9/explant, its length was 11.3 ㎝, and its fresh weight was 2.5 g. When the bulblets were cultured in MS medium with 2 ㎎/L kinetin and 0.5 ㎎/L NAA, the addition of 30 ㎎/L adenine improved their proliferation and growth significantly, the highest bulblet formation rate (48%) was obtained. The addition of 7% sucrose also increased the bulblet formation rate at the highest frequency of 98.2%. The shoots were shown be more vigorously proliferated at the secondary subculture stage rather than primary culture stage, their propagation rate was 80% after subculture.