Fetal Bovine Serum (FBS), which offers nutrients for cell growth, has been the most widely used serum in proliferation studies. It does, however, necessitate screening because it contains unidentified elements, as well as viruses and prions, which pose a risk of infection. Apis mellifera royal jelly has the potential to replace FBS as a serum. In this study, a comparison of the effectiveness of three royal jelly treatments on lymphocyte cell proliferation will be performed: untreated royal jelly, soluble royal jelly, and hydrolyzed royal jelly, which is then freeze-dyed to generate a powdered product. Lymphocyte cells were cultured with various concentrations (2.5%, 5%, 7.5%, and 10%) and three different treatments of royal jelly. The results of these various concentrations were continued for up to 48 hours with continuous checking every 24 hours measured by the Microtetrazolium (MTT) assay. According to the results, lymphocyte cells cultured with the addition of a 2.5% concentration of untreated royal jelly Apis mellifera had significant differences, with the percent cell viability of 77.15% at 24 hours and 58.44% at 48 hours. This value was higher than that of soluble royal jelly (64.13% and 38.52%) and royal jelly hydrolyzate (71.08% and 54.59%) with the addition of the same concentration. © 2024 American Institute of Physics Inc.. All rights reserved.