<p><strong>Background. </strong>Sugarcane (<em>Saccharum officinarum</em> L.) is one of the important commodities in Indonesia as raw material for sugar production. The constraints for sugarcane cultivation are limited availability and poor quality of seedling. The utilization of liquid culture in tissue culture enables to produce good quality seedlings, free of viruses and diseases, and can generate a large quantity in a short time. Liquid culture can produce embryogenic cells faster since embryogenic callus can develop easily in liquid media. <strong>Objective</strong>. This study determined the appropriate pH and concentration of sucrose in the liquid culture of embryogenic cells. <strong>Methodology</strong>. This study was conducted in 3 stages of in vitro culture, which were callus induction, solid media proliferation, and liquid media proliferation that using pH and sucrose treatments. <strong>Result</strong>. The results showed that pH 6.5 + 3% sucrose (A3B1) produced the highest number of callus with the average callus on the first week 29 callus, the second week 8,33 callus and the third week 4,33 callus and the color did not easily turn to brown with color 5Y 8/2. <strong>Implications</strong>. The protocol was developed that allows embryogenic callus of sugarcane plants to be obtained in liquid media that can be used for embryogenic callus proliferation. <strong>Conclusions</strong>. pH 6.5 + sucrose 3% (A3B1) was the best combination treatment of pH and sucrose for sugarcane callus proliferation in liquid culture media, which produced the highest number of calluses and color of callus that not easily turned brown.</p>