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In vitro system for the growth and asexual multiplication of Taenia crassiceps cysticerci

Published online by Cambridge University Press:  27 September 2022

Lucía Celia Abril García ,
Matías Gastón Pérez ,
María Eugenia Ancarola ,
Mara Cecilia Rosenzvit  and
Marcela Alejandra Cucher Open the ORCID record for Marcela Alejandra Cucher [Opens in a new window]
Lucía Celia Abril García
Affiliation:
Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina Institute of Research on Microbiology and Medical Parasitology (IMPaM, UBA-CONICET), University of Buenos Aires, Buenos Aires, Argentina
Matías Gastón Pérez
Affiliation:
Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina Institute of Research on Microbiology and Medical Parasitology (IMPaM, UBA-CONICET), University of Buenos Aires, Buenos Aires, Argentina
María Eugenia Ancarola
Affiliation:
Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina Institute of Research on Microbiology and Medical Parasitology (IMPaM, UBA-CONICET), University of Buenos Aires, Buenos Aires, Argentina
Mara Cecilia Rosenzvit
Affiliation:
Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina Institute of Research on Microbiology and Medical Parasitology (IMPaM, UBA-CONICET), University of Buenos Aires, Buenos Aires, Argentina
Marcela Alejandra Cucher*
Affiliation:
Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina Institute of Research on Microbiology and Medical Parasitology (IMPaM, UBA-CONICET), University of Buenos Aires, Buenos Aires, Argentina
*
Author for correspondence: Marcela Alejandra Cucher, E-mail: [email protected]
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Abstract

Taenia solium is the aetiological agent of cysticercosis, a zoonosis that causes severe health and economic losses across Latin America, Africa and Asia. The most serious manifestation of the disease is neurocysticercosis, which occurs when the larval stage (cysticercus) establishes in the central nervous system. Using Taenia crassiceps as an experimental model organism for the study of cysticercosis, we aimed to identify the in vitro conditions necessary to allow parasite development at the short- and long terms. First, cysticerci were incubated for 15 days in different media and parasite densities. The number of buddings and cysticerci diameter were measured to evaluate asexual multiplication and parasite growth, respectively. Vitality was determined by trypan blue staining and morphology analysis. As a result, high cysticerci density and medium containing FBS and the excretion/secretion (E/S) products of feeder cells induced parasite survival, growth and multiplication. Then, the long-term (5 weeks) incubation of the parasites in co-culture with feeder cells was evaluated. Consequently, the mammalian cell lines induced a significant increase in total parasite volume while axenic cultures did not show any statistically significant change over time. In this study, the proper conditions to maintain T. crassiceps in vitro are described for the first time in a simpler and more controlled setting other than experimental infections. In addition, it was shown that cysticerci growth, survival and asexual multiplication depend on a complex network of secreted factors from both parasite and host.

Keywords

Cysticercushelminthin vitrometacestodemultiplicationTaenia
Type
Research Article
Information
Parasitology , Volume 149 , Issue 13 , November 2022 , pp. 1775 - 1780
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Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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References

Adalid-Peralta, L, Rosas, G, Arce-Sillas, A, Bobes, RJ, Cárdenas, G, Hernández, M, Trejo, C, Meneses, G, Hernández, B, Estrada, K, Fleury, A, Laclette, JP, Larralde, C, Sciutto, E and Fragoso, G (2017) Effect of transforming growth factor-β upon Taenia solium and Taenia crassiceps cysticerci. Scientific Reports 7, 12345.10.1038/s41598-017-12202-zCrossRefGoogle ScholarPubMed
Ancarola, ME, Marcilla, A, Herz, M, Macchiaroli, N, Pérez, M, Asurmendi, S, Brehm, K, Poncini, C, Rosenzvit, M and Cucher, M (2017) Cestode parasites release extracellular vesicles with microRNAs and immunodiagnostic proteins cargo. International Journal for Parasitology 47, 675–686.10.1016/j.ijpara.2017.05.003CrossRefGoogle ScholarPubMed
Colombo, M, Raposo, G and Théry, C (2014) Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annual Review of Cell and Developmental Biology 30, 255–289.CrossRefGoogle ScholarPubMed
Coral-Almeida, M, Gabriël, S, Abatih, EN, Praet, N, Benitez, W and Dorny, P (2015) Taenia solium human cysticercosis: a systematic review of sero-epidemiological data from endemic zones around the world. PLoS Neglected Tropical Diseases 9, e0003919.10.1371/journal.pntd.0003919CrossRefGoogle ScholarPubMed
Del Brutto, OH (2012) Neurocysticercosis: a review. The Scientific World Journal 2012, Article ID 159821. doi: 10.1100/2012/159821CrossRefGoogle ScholarPubMed
Díaz-Orea, MA, de Aluja, AS, de L'Erosa, M, Gomez-Conde, E, Castellanos Sánchez, VO, Willms, K, Sciutto, E and Fragoso, G (2007) Different effects of chorionic gonadotropin on Taenia crassiceps and Taenia solium cysticerci cultured in vitro. Journal of Parasitology 93, 1518–1520.CrossRefGoogle ScholarPubMed
Escobedo, G, Larralde, C, Chavarria, A, Cerbón, MA and Morales-Montor, J (2004) Molecular mechanisms involved in the differential effects of sex steroids on the reproduction and infectivity of Taenia crassiceps. Journal of Parasitology 90, 1235–1244.10.1645/GE-297RCrossRefGoogle ScholarPubMed
Escobedo, G, Romano, MC and Morales-Montor, J (2009) Differential in vitro effects of insulin on Taenia crassiceps and Taenia solium cysticerci. Journal of Helminthology 83, 403–412.CrossRefGoogle ScholarPubMed
Escobedo, G, Soldevila, G, Ortega-Pierres, G, Chávez-Ríos, JR, Nava, K, Fonseca-Liñán, R, López-Griego, L, Hallal-Calleros, C, Ostoa-Saloma, P and Morales-Montor, J (2010) A new MAP kinase protein involved in estradiol-stimulated reproduction of the helminth parasite Taenia crassiceps. Journal of Biomedicine and Biotechnology 2010, Article ID 747121. doi: 10.1155/2010/747121CrossRefGoogle ScholarPubMed
Esquivel Velázquez, M, Hernández, R, Larralde, C and Ostoa Saloma, P (2014) Crosstalk among Taenia crassiceps (ORF strain) cysts regulates their rates of budding by ways of soluble and contact signals exchanged between them. BioMed Research International 2014, Article ID 703693. doi: 10.1155/2014/703693Google Scholar
Galán-Puchades, MT, Yang, Y, Marcilla, A, Choe, S, Park, H, Osuna, A and Eom, KS (2016) First ultrastructural data on the human tapeworm Taenia asiatica eggs by scanning and transmission electron microscopy (SEM, TEM). Parasitology Research 115, 3649–3655. doi: 10.1007/s00436-016-5165-4CrossRefGoogle Scholar
GBD 2017 Disease and Injury Incidence and Prevalence Collaborators (2018) Global, regional, and national incidence, prevalence, and years lived with disability for 354 diseases and injuries for 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet (London, England) 392, 1789–1858.10.1016/S0140-6736(18)32279-7CrossRefGoogle Scholar
Hemphill, A and Gottstein, B (1995) Immunology and morphology studies on the proliferation of in vitro cultivated Echinococcus multilocularis metacestodes. Parasitology Research 81, 605–614.10.1007/BF00932028CrossRefGoogle ScholarPubMed
Jura, H, Bader, A, Hartmann, M, Maschek, H and Frosch, M (1996) Hepatic tissue culture model for study of host-parasite interactions in alveolar echinococcosis. Infection and Immunity 64, 3484–3490.10.1128/iai.64.9.3484-3490.1996CrossRefGoogle ScholarPubMed
Liang, P, Mao, L, Zhang, S, Guo, X, Liu, G, Wang, L, Hou, J, Zheng, Y and Luo, X (2019) Identification and molecular characterization of exosome-like vesicles derived from the Taenia asiatica adult worm. Acta Tropica 198, 105036.10.1016/j.actatropica.2019.05.027CrossRefGoogle ScholarPubMed
McSorley, HJ and Maizels, RM (2012) Helminth infections and host immune regulation. Clinical Microbiology Reviews 25, 585–608.10.1128/CMR.05040-11CrossRefGoogle ScholarPubMed
Ostoa-Saloma, P, Ostoa-Jacobo, P, Esquivel-Velázquez, M, Bazúa, S and Larralde, C (2010) Budding of Taenia crassiceps cysticerci in vitro is promoted by crowding in addition to hormonal, stress, and energy-related signals. Journal of Biomedicine and Biotechnology 2010, Article ID 510978. doi: 10.1155/2010/510978CrossRefGoogle ScholarPubMed
Sánchez-López, CM, Trelis, M, Bernal, D and Marcilla, A (2021) Overview of the interaction of helminth extracellular vesicles with the host and their potential functions and biological applications. Molecular Immunology 134, 228–235.CrossRefGoogle ScholarPubMed
Sotillo, J, Robinson, MW, Kimber, MJ, Cucher, M, Ancarola, ME, Nejsum, P, Marcilla, A, Eichenberger, RM and Tritten, L (2020) The protein and microRNA cargo of extracellular vesicles from parasitic helminths – current status and research priorities. International Journal for Parasitology 50, 635–645.10.1016/j.ijpara.2020.04.010CrossRefGoogle ScholarPubMed
Spiliotis, M and Brehm, K (2009) Axenic in vitro cultivation of Echinococcus multilocularis metacestode vesicles and the generation of primary cell cultures. Methods in Molecular Biology (Clifton, N.J.) 470, 245–262.CrossRefGoogle ScholarPubMed
Spiliotis, M, Tappe, D, Sesterhenn, L and Brehm, K (2004) Long-term in vitro cultivation of Echinococcus multilocularis metacestodes under axenic conditions. Parasitology Research 92, 430–432.10.1007/s00436-003-1046-8CrossRefGoogle ScholarPubMed
Spiliotis, M, Lechner, S, Tappe, D, Scheller, C, Krohne, G and Brehm, K (2008) Transient transfection of Echinococcus multilocularis primary cells and complete in vitro regeneration of metacestode vesicles. International Journal for Parasitology 38, 1025–1039.10.1016/j.ijpara.2007.11.002CrossRefGoogle ScholarPubMed
Théry, C, Amigorena, S, Raposo, G and Clayton, A (2006) Isolation and characterization of exosomes from cell culture supernatants. Current Protocols in Cell Biology 3, 3.22.1–3.22.29.Google ScholarPubMed
van Niel, G, D'Angelo, G and Raposo, G (2018) Shedding light on the cell biology of extracellular vesicles. Nature Reviews Molecular Cell Biology 19, 213–228.10.1038/nrm.2017.125CrossRefGoogle ScholarPubMed
Wang, L-Q, Liu, T-L, Liang, P-H, Zhang, S-H, Li, T-S, Li, Y-P, Liu, G-X, Mao, L and Luo, X-N (2020) Characterization of exosome-like vesicles derived from Taenia pisiformis cysticercus and their immunoregulatory role on macrophages. Parasites & Vectors 13, 318.CrossRefGoogle ScholarPubMed
Willms, K and Zurabian, R (2010) Taenia crassiceps: in vivo and in vitro models. Parasitology 137, 335–346. doi: 10.1017/S0031182009991442CrossRefGoogle ScholarPubMed
World Health Organization (2020) Ending the neglect to attain the Sustainable Development Goals: a road map for neglected tropical diseases 2021–2030.Google Scholar