Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model
<i>Toxoplasma gondii</i> is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of <i>T. gondii</i> comes from studies relying on the RH strain propagate...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Pathogens |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-0817/11/9/1038 |
| _version_ | 1797483786009575424 |
|---|---|
| author | Maria João Gargaté Anabela Vilares Idalina Ferreira Tânia Reis Susana Martins Joana Mendonça Vítor Borges João Paulo Gomes |
| author_facet | Maria João Gargaté Anabela Vilares Idalina Ferreira Tânia Reis Susana Martins Joana Mendonça Vítor Borges João Paulo Gomes |
| author_sort | Maria João Gargaté |
| collection | DOAJ |
| description | <i>Toxoplasma gondii</i> is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of <i>T. gondii</i> comes from studies relying on the RH strain propagated in mice. According to the recommendations concerning the animal welfare, we assayed in vitro/in vivo procedures to replace, or at least reduce, the demanding animal model for strain propagation. We evaluated the genetic and phenotypic stability of the RH strain throughout its parallel continuous propagation in mice, in human foreskin fibroblasts (HFF) and in an alternate fashion of these two procedures. We also assessed the virulence impact on the RH strain after different periods of its long-term propagation strictly in cells. The RH strain completely lost its virulence after long-term passage in HFF. Nevertheless, we obtained a successful outcome with the alternate passaging of the parasite in HFF and in mice as this approach enabled <i>T. gondii</i> to maintain the evaluated phenotypic properties, mainly its virulence potential. Also, no genetic changes were observed in genes known to be highly polymorphic or involved in pathoadaptation. In conclusion, the alternate model seems to be a feasible method for <i>T. gondii</i> propagation and maintenance, strongly impacting the number of sacrificed mice. |
| first_indexed | 2024-03-09T22:53:00Z |
| format | Article |
| id | doaj.art-2ecb2cc0737149d7a8b97583963ef379 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-09T22:53:00Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-2ecb2cc0737149d7a8b97583963ef3792023-11-23T18:16:23ZengMDPI AGPathogens2076-08172022-09-01119103810.3390/pathogens11091038Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice ModelMaria João Gargaté0Anabela Vilares1Idalina Ferreira2Tânia Reis3Susana Martins4Joana Mendonça5Vítor Borges6João Paulo Gomes7National Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalNational Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalNational Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalNational Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalNational Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalInnovation and Technology Unit, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalGenomics and Bioinformatics Unit, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, PortugalGenomics and Bioinformatics Unit, Department of Infectious Diseases, National Institute of Health Doctor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisbon, Portugal<i>Toxoplasma gondii</i> is an obligate intracellular protozoan. In pregnant women, it can lead to severe birth defects or intrauterine death of the fetus. Most of what is currently know on cell biology of <i>T. gondii</i> comes from studies relying on the RH strain propagated in mice. According to the recommendations concerning the animal welfare, we assayed in vitro/in vivo procedures to replace, or at least reduce, the demanding animal model for strain propagation. We evaluated the genetic and phenotypic stability of the RH strain throughout its parallel continuous propagation in mice, in human foreskin fibroblasts (HFF) and in an alternate fashion of these two procedures. We also assessed the virulence impact on the RH strain after different periods of its long-term propagation strictly in cells. The RH strain completely lost its virulence after long-term passage in HFF. Nevertheless, we obtained a successful outcome with the alternate passaging of the parasite in HFF and in mice as this approach enabled <i>T. gondii</i> to maintain the evaluated phenotypic properties, mainly its virulence potential. Also, no genetic changes were observed in genes known to be highly polymorphic or involved in pathoadaptation. In conclusion, the alternate model seems to be a feasible method for <i>T. gondii</i> propagation and maintenance, strongly impacting the number of sacrificed mice.https://www.mdpi.com/2076-0817/11/9/1038<i>Toxoplasma gondii</i>3Rsstrain propagationmice modelin vitro propagationvirulence |
| spellingShingle | Maria João Gargaté Anabela Vilares Idalina Ferreira Tânia Reis Susana Martins Joana Mendonça Vítor Borges João Paulo Gomes Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model Pathogens <i>Toxoplasma gondii</i> 3Rs strain propagation mice model in vitro propagation virulence |
| title | Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model |
| title_full | Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model |
| title_fullStr | Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model |
| title_full_unstemmed | Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model |
| title_short | Parallel Propagation of <i>Toxoplasma gondii</i> In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model |
| title_sort | parallel propagation of i toxoplasma gondii i in vivo in vitro and in alternate model towards less dependence on the mice model |
| topic | <i>Toxoplasma gondii</i> 3Rs strain propagation mice model in vitro propagation virulence |
| url | https://www.mdpi.com/2076-0817/11/9/1038 |
| work_keys_str_mv | AT mariajoaogargate parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT anabelavilares parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT idalinaferreira parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT taniareis parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT susanamartins parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT joanamendonca parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT vitorborges parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel AT joaopaulogomes parallelpropagationofitoxoplasmagondiiiinvivoinvitroandinalternatemodeltowardslessdependenceonthemicemodel |