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Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum

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 poss...

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Autores principales: Habibzadeh, Sima, Doroud, Delaram, Taheri, Tahereh, Seyed, Negar, Rafati, Sima
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Pasteur Institute of Iran 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8487682/
https://www.ncbi.nlm.nih.gov/pubmed/34481426
http://dx.doi.org/10.52547/ibj.25.5.349
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author Habibzadeh, Sima
Doroud, Delaram
Taheri, Tahereh
Seyed, Negar
Rafati, Sima
author_facet Habibzadeh, Sima
Doroud, Delaram
Taheri, Tahereh
Seyed, Negar
Rafati, Sima
author_sort Habibzadeh, Sima
collection PubMed
description 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 (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.
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spelling pubmed-84876822021-10-13 Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum Habibzadeh, Sima Doroud, Delaram Taheri, Tahereh Seyed, Negar Rafati, Sima Iran Biomed J Full Length 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 (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. Pasteur Institute of Iran 2021-09 2021-08-29 /pmc/articles/PMC8487682/ /pubmed/34481426 http://dx.doi.org/10.52547/ibj.25.5.349 Text en https://creativecommons.org/licenses/by/3.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License, (http://creativecommons.org/licenses/by/3.0/ (https://creativecommons.org/licenses/by/3.0/) ) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Full Length
Habibzadeh, Sima
Doroud, Delaram
Taheri, Tahereh
Seyed, Negar
Rafati, Sima
Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title_full Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title_fullStr Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title_full_unstemmed Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title_short Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum
title_sort leishmania parasite: the impact of new serum-free medium as an alternative for fetal bovine serum
topic Full Length
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8487682/
https://www.ncbi.nlm.nih.gov/pubmed/34481426
http://dx.doi.org/10.52547/ibj.25.5.349
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