en Molecular Immunology & Vaccines Molecular Immunology & Vaccines مقاله کامل Full Length/Original Article <strong>Background: </strong>Flagellated protozoan of the genus <em>Leishmania</em> is the causative agent of vector-borne parasitic diseases of leishmaniasis. Since the production of recombinant pharmaceutical proteins requires the cultivation of host cells in a serum-free medium, the elimination of FBS can improve the possibility of large-scale culture of <em>Leishmania </em>parasite. In the current study, we aimed at evaluating a new serum-free medium in <em>Leishmania </em>parasite culture for future live <em>Leishmania</em> vaccine purposes. <strong>Methods: </strong>Recombinant <em>L. tarentolae</em> secreting PpSP15-EGFP and wild type <em>L. major</em> were cultured in serum-free (complete serum-free medium [CSFM]) and serum-supplemented medium. The growth rate, protein expression, and infectivity of cultured parasites in both conditions was then evaluated and compared. <strong>Results: </strong>Diff-Quick staining and epi-fluores&shy;cence microscopy examination displayed the typical morphology of<em> L. major </em>and<em> L. tarentolae-</em>PpSP15-EGFP promastigote grown in CSFM medium. The amount of EGFP expression was similar in CSMF medium compared to M199 supplemented with 5% FBS in flow cytometry analysis of <em>L. tarentolae</em>-PpSP15-EGFP parasite. Also, a similar profile of PpSP15-EGFP proteins was recognized in Western blot analysis of <em>L. tarentolae-</em>PpSP15-EGFP cultured in CSMF and the serum-supplemented medium. Footpad swelling and parasite load measurements showed the ability of CSFM medium to support the <em>L. major</em> infectivity in BALB/C mice.<strong> Conclusion: </strong>This study demonstrated that CSFM can be a promising substitute for FBS supplemented medium in parasite culture for live vaccination purposes.<br> <gdiv></gdiv><gdiv></gdiv> Growth rate, L. major, L. tarentolae, Serum-free medium, PpSP15-EGFP protein 349 358 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-4430-1&slc_lang=en&sid=1 Sima Habibzadeh Simahabibzade@gmail.com 0000-0001-8036-8419 No Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Delaram Doroud d_doroud@yahoo.com 0000-0003-4167-9921 No Quality Control Department, Production and Research Complex, Pasteur institute of Iran, Tehran, Iran Tahereh Taheri tahereh_t@yahoo.com 0000-0002-2619-6879 No Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran Negar Seyed negarse@gmail.com 0000-0001-8249-6692 No Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran Sima Rafati s_rafati@yahoo.com 0000-0002-7221-1320 Yes Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran