Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes
Leishmania (Mundinia) orientalis is a human pathogen causing leishmaniasis and studies on the properties of metacyclic promastigotes, the parasite’s infective stage, are required for a better understanding of its transmission and infection. However, information on cultivation for mass production of...
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.992741/full |
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| author | Wetpisit Chanmol Narissara Jariyapan Kanok Preativatanyou Chonlada Mano Pongsri Tippawangkosol Pongsri Tippawangkosol Pradya Somboon Paul A. Bates |
| author_facet | Wetpisit Chanmol Narissara Jariyapan Kanok Preativatanyou Chonlada Mano Pongsri Tippawangkosol Pongsri Tippawangkosol Pradya Somboon Paul A. Bates |
| author_sort | Wetpisit Chanmol |
| collection | DOAJ |
| description | Leishmania (Mundinia) orientalis is a human pathogen causing leishmaniasis and studies on the properties of metacyclic promastigotes, the parasite’s infective stage, are required for a better understanding of its transmission and infection. However, information on cultivation for mass production of L. orientalis metacyclic promastigotes and factors that stimulate their metacyclogenesis is limited. Therefore, the objective of this study was to develop a suitable methodology for generating promastigote cultures containing a high proportion and number of L. orientalis metacyclic promastigotes. Various media, i.e., Schneider’s insect medium, Medium 199 and Grace’s insect medium, supplemented with various quantities of dithiothreitol, Basal Medium Eagle vitamins, pooled human urine, and fetal bovine serum, were optimized for metacyclogenesis. The results revealed that the optimum culture medium and conditions of those tested were Schneider’s insect medium supplemented with 100 μM dithiothreitol, 1% (v/v) Basal Medium Eagle vitamins, 2% (v/v) pooled human urine, and 10% (v/v) fetal bovine serum, pH 5.0 at 26°C. We also demonstrated that L. orientalis metacyclic promastigotes could be purified and enriched by negative selection using peanut lectin. Under these culture conditions, the highest yield of metacyclic promastigotes was obtained with a significantly higher percentage of parasite survival, resistance to complement-mediated lysis, and infection index in THP-1 macrophage cells compared to parasites cultured without media supplements at neutral pH. This is the first report providing a reliable method for mass production of L. orientalis metacyclic promastigotes for in vivo infections and other experimental studies of this emerging parasite in the future. |
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| id | doaj.art-3866de2c122e42b88d1e1b39f610c265 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-13T06:01:42Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-3866de2c122e42b88d1e1b39f610c2652022-12-22T02:59:25ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-09-011210.3389/fcimb.2022.992741992741Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotesWetpisit Chanmol0Narissara Jariyapan1Kanok Preativatanyou2Chonlada Mano3Pongsri Tippawangkosol4Pongsri Tippawangkosol5Pradya Somboon6Paul A. Bates7School of Allied Health Sciences, Walailak University, Nakhonsithammarat, ThailandCenter of Excellence in Vector Biology and Vector-Borne Disease, Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandCenter of Excellence in Vector Biology and Vector-Borne Disease, Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandFaculty of Medicine, Chiang Mai University, Chiang Mai, ThailandSchool of Allied Health Sciences, Walailak University, Nakhonsithammarat, ThailandDepartment of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai, ThailandDepartment of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai, ThailandDivision of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster, United KingdomLeishmania (Mundinia) orientalis is a human pathogen causing leishmaniasis and studies on the properties of metacyclic promastigotes, the parasite’s infective stage, are required for a better understanding of its transmission and infection. However, information on cultivation for mass production of L. orientalis metacyclic promastigotes and factors that stimulate their metacyclogenesis is limited. Therefore, the objective of this study was to develop a suitable methodology for generating promastigote cultures containing a high proportion and number of L. orientalis metacyclic promastigotes. Various media, i.e., Schneider’s insect medium, Medium 199 and Grace’s insect medium, supplemented with various quantities of dithiothreitol, Basal Medium Eagle vitamins, pooled human urine, and fetal bovine serum, were optimized for metacyclogenesis. The results revealed that the optimum culture medium and conditions of those tested were Schneider’s insect medium supplemented with 100 μM dithiothreitol, 1% (v/v) Basal Medium Eagle vitamins, 2% (v/v) pooled human urine, and 10% (v/v) fetal bovine serum, pH 5.0 at 26°C. We also demonstrated that L. orientalis metacyclic promastigotes could be purified and enriched by negative selection using peanut lectin. Under these culture conditions, the highest yield of metacyclic promastigotes was obtained with a significantly higher percentage of parasite survival, resistance to complement-mediated lysis, and infection index in THP-1 macrophage cells compared to parasites cultured without media supplements at neutral pH. This is the first report providing a reliable method for mass production of L. orientalis metacyclic promastigotes for in vivo infections and other experimental studies of this emerging parasite in the future.https://www.frontiersin.org/articles/10.3389/fcimb.2022.992741/fullLeishmaniaLeishmania orientalisleishmaniasismetacyclogenesismetacyclic promastigotestimulation |
| spellingShingle | Wetpisit Chanmol Narissara Jariyapan Kanok Preativatanyou Chonlada Mano Pongsri Tippawangkosol Pongsri Tippawangkosol Pradya Somboon Paul A. Bates Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes Frontiers in Cellular and Infection Microbiology Leishmania Leishmania orientalis leishmaniasis metacyclogenesis metacyclic promastigote stimulation |
| title | Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes |
| title_full | Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes |
| title_fullStr | Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes |
| title_full_unstemmed | Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes |
| title_short | Stimulation of metacyclogenesis in Leishmania (Mundinia) orientalis for mass production of metacyclic promastigotes |
| title_sort | stimulation of metacyclogenesis in leishmania mundinia orientalis for mass production of metacyclic promastigotes |
| topic | Leishmania Leishmania orientalis leishmaniasis metacyclogenesis metacyclic promastigote stimulation |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.992741/full |
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