Volume 25, Issue 5 (9-2021)                   IBJ 2021, 25(5): 349-358 | Back to browse issues page

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Habibzadeh S, Doroud D, Taheri T, Seyed N, Rafati S. Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum. IBJ 2021; 25 (5) :349-358
URL: http://ibj.pasteur.ac.ir/article-1-3410-en.html
Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
Sima Habibzadeh , Delaram Doroud , Tahereh Taheri , Negar Seyed , Sima Rafati *
Abstract:  
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.
Keywords: Growth rate, L. major, L. tarentolae, Serum-free medium, PpSP15-EGFP protein
Full-Text [PDF 1098 kb]      
Type of Study: Full Length/Original Article | Subject: Molecular Immunology & Vaccines
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