Abstract: [Objective] The research aimed to explore the preparation method of chromosome of cell lines（MDEC-07114） in Musca domestica embryo. [Method] The chromosome was prepared with MDEC-07114 cells as materials. The experimental groups were set up as follows：groups of 5 and 10 mg/L colchicine;groups of treating with colchicine for 5,6 and 10 h,pre-fixative group and the fixed group. [Result] More cell division and good chromosome morphology can be observed in the group of treating with 10 mg/L colchicine for 5 h. And clear and separated chromosome were observed in pre-fixative group. [Conclusion] The prepared chromosome of MDEC-07114 cells after treating with 10 mg/L colchicine for 5 h and pre-fixative treatment had the best effects.