Abstract:The cotyledon protoplasts were isolated from sterile seedling of Xinjiang muskmelon (Cucumis melo L.) and cultured in modified Miller medium. The high frequency division of the regenerated cells was observed It was indicated that the agarose bead culture with B6S3 nurse cells is the most suitable for the cotyledon protoplast of Xinjiang muskmelon when compared with thin liquid culture and double layer culture. The intact plants were differentiated from the regenerated calli by two steps of culture with liquid first and then solid medium.