TY - JOUR JF - انستیتو-پاستور-ایران JO - IBJ VL - 25 IS - 5 PY - 2021 Y1 - 2021/9/01 TI - Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum TT - N2 - Background: Flagellated protozoan of the genus Leishmania 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 Leishmania parasite. In the current study, we aimed at evaluating a new serum-free medium in Leishmania parasite culture for future live Leishmania vaccine purposes. Methods: Recombinant L. tarentolae secreting PpSP15-EGFP and wild type L. major 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. Results: Diff-Quick staining and epi-fluores­cence microscopy examination displayed the typical morphology of L. major and L. tarentolae-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 L. tarentolae-PpSP15-EGFP parasite. Also, a similar profile of PpSP15-EGFP proteins was recognized in Western blot analysis of L. tarentolae-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 L. major infectivity in BALB/C mice. Conclusion: This study demonstrated that CSFM can be a promising substitute for FBS supplemented medium in parasite culture for live vaccination purposes. SP - 349 EP - 358 AU - Habibzadeh, Sima AU - Doroud, Delaram AU - Taheri, Tahereh AU - Seyed, Negar AU - Rafati, Sima AD - Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran KW - Growth rate KW - L. major KW - L. tarentolae KW - Serum-free medium KW - PpSP15-EGFP protein UR - http://ibj.pasteur.ac.ir/article-1-3410-en.html DO - 10.52547/ibj.25.5.349 ER -