Abstract: The establishment of an effective and reliable cell suspension culture method is of great significance for protoplast culture and transgenic research. With shoot tips and immature inflorescences as explants, we carried out research on the establishment of sorghum cell suspension lines by the three ways of callus shake culture, homogenate collection culture and whole explants direct culture. The results indicate that the immature inflorescence is the better explants for cell suspension culture of sorghum, compared with shoot tips. All the three culture methods of immature inflorescences are applicable. Among them, callus shake culture is the most stable and reliable method. Although the other two methods have less stability and effectiveness, but they enjoy better comprehensive cell growth status including higher cell propagation capability. The results also show that the shoot tip callus shake culture is also a reliable preparation way of cell suspensions, the shoot tips homogenate culture is less reliable, while whole explants direct culture gave poor result and should not be used.