A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites
Abstract This study describes a simple method for the large‐scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were colle...
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| Language: | English |
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Wiley
2021-03-01
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| Series: | Veterinary Medicine and Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/vms3.364 |
| _version_ | 1818611196106899456 |
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| author | Saeed El‐Ashram Yu Zhang Yongsheng Ji Dina Salama Kun Mei Li Zhili Huang Shujian Haoji Zhang Shawky M. Aboelhadid Reem A. Alajmi Dina M. Metwally Manal F. El‐Khadragy Billy M. Hargis Guillermo Tellez‐Isaias Beniamino T. Cenci‐Goga Musafiri Karama Munyaradzi C. Marufu Fathi Abouhajer Gamal Ali Abdelhafez Hamady Abeer El Wakil Ibrahim Al Nasr Xun Suo |
| author_facet | Saeed El‐Ashram Yu Zhang Yongsheng Ji Dina Salama Kun Mei Li Zhili Huang Shujian Haoji Zhang Shawky M. Aboelhadid Reem A. Alajmi Dina M. Metwally Manal F. El‐Khadragy Billy M. Hargis Guillermo Tellez‐Isaias Beniamino T. Cenci‐Goga Musafiri Karama Munyaradzi C. Marufu Fathi Abouhajer Gamal Ali Abdelhafez Hamady Abeer El Wakil Ibrahim Al Nasr Xun Suo |
| author_sort | Saeed El‐Ashram |
| collection | DOAJ |
| description | Abstract This study describes a simple method for the large‐scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were collected from chronically infected mice. Harvested cells and brain tissues were incubated in Hanks balanced salt solution (HBSS), containing 0.25% trypsin and 0.5% taurodeoxycholic acid (TDC) for 5 min. Subsequent washes in phosphate buffered saline (PBS) were conducted, and the cell viability of the preparations was good, as determined by flow cytometry and ability to reinfect HFF cells and propagate in mice. The purification procedure allowed for a rapid preparation of pure T. gondii tachyzoites and bradyzoites in sufficient quantity that can be used for downstream procedures. The advantage of the new method is that it is convenient and inexpensive. |
| first_indexed | 2024-12-16T15:26:29Z |
| format | Article |
| id | doaj.art-5c5de8061d014bf398728939b7146545 |
| institution | Directory Open Access Journal |
| issn | 2053-1095 |
| language | English |
| last_indexed | 2024-12-16T15:26:29Z |
| publishDate | 2021-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine and Science |
| spelling | doaj.art-5c5de8061d014bf398728939b71465452022-12-21T22:26:29ZengWileyVeterinary Medicine and Science2053-10952021-03-017235736110.1002/vms3.364A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoitesSaeed El‐Ashram0Yu Zhang1Yongsheng Ji2Dina Salama3Kun Mei4Li Zhili5Huang Shujian6Haoji Zhang7Shawky M. Aboelhadid8Reem A. Alajmi9Dina M. Metwally10Manal F. El‐Khadragy11Billy M. Hargis12Guillermo Tellez‐Isaias13Beniamino T. Cenci‐Goga14Musafiri Karama15Munyaradzi C. Marufu16Fathi Abouhajer17Gamal Ali Abdelhafez Hamady18Abeer El Wakil19Ibrahim Al Nasr20Xun Suo21College of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaCollege of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaSchool of Basic Medical Sciences Division of Life Science and Medicine University of Science and Technology of China Hefei ChinaDepartment of Parasitology and Animal Disease National Research Centre Dokki Giza EgyptCollege of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaCollege of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaCollege of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaCollege of Life Science and Engineering Foshan University Foshan Guangdong Province ChinaParasitology Department Faculty of Veterinary Medicine Beni Suef University Beni‐Suef EgyptDepartment of Zoology Faculty of Science King Saud University Riyadh Saudi ArabiaDepartment of Zoology Faculty of Science King Saud University Riyadh Saudi ArabiaDepartment of Zoology and Entomology Faculty of Science Helwan University Cairo EgyptDepartment of Poultry Science University of Arkansas Fayetteville AR USADepartment of Poultry Science University of Arkansas Fayetteville AR USADipartimento di Medicina Veterinaria Laboratorio di Ispezione degli Alimenti di Origine Animale University of Perugia Perugia ItalyDepartment of Paraclinical Sciences Faculty of Veterinary Science University of Pretoria Onderstepoort South AfricaDepartment of Veterinary Tropical Diseases Faculty of Veterinary Science University of Pretoria Onderstepoort South AfricaFaculty of Education Asmarya University for Islamic Sciences Zliten LibyaDepartment of Animal Production Faculty of Agriculture Al‐Azhar University Nasr City, Cairo EgyptDepartment of Biological & Geological Sciences Alexandria University Alexandria EgyptCollege of Science and Arts in Unaizah Qassim University Unaizah Saudi ArabiaNational Animal Protozoa Laboratory & College of Veterinary Medicine China Agricultural University Beijing ChinaAbstract This study describes a simple method for the large‐scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were collected from chronically infected mice. Harvested cells and brain tissues were incubated in Hanks balanced salt solution (HBSS), containing 0.25% trypsin and 0.5% taurodeoxycholic acid (TDC) for 5 min. Subsequent washes in phosphate buffered saline (PBS) were conducted, and the cell viability of the preparations was good, as determined by flow cytometry and ability to reinfect HFF cells and propagate in mice. The purification procedure allowed for a rapid preparation of pure T. gondii tachyzoites and bradyzoites in sufficient quantity that can be used for downstream procedures. The advantage of the new method is that it is convenient and inexpensive.https://doi.org/10.1002/vms3.364flow cytometrypurificationT. gondii tachyzoites and bradyzoitestrypsintaurodeoxycholic acid |
| spellingShingle | Saeed El‐Ashram Yu Zhang Yongsheng Ji Dina Salama Kun Mei Li Zhili Huang Shujian Haoji Zhang Shawky M. Aboelhadid Reem A. Alajmi Dina M. Metwally Manal F. El‐Khadragy Billy M. Hargis Guillermo Tellez‐Isaias Beniamino T. Cenci‐Goga Musafiri Karama Munyaradzi C. Marufu Fathi Abouhajer Gamal Ali Abdelhafez Hamady Abeer El Wakil Ibrahim Al Nasr Xun Suo A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites Veterinary Medicine and Science flow cytometry purification T. gondii tachyzoites and bradyzoites trypsin taurodeoxycholic acid |
| title | A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites |
| title_full | A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites |
| title_fullStr | A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites |
| title_full_unstemmed | A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites |
| title_short | A rapid and simple single‐step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites |
| title_sort | rapid and simple single step method for the purification of toxoplasma gondii tachyzoites and bradyzoites |
| topic | flow cytometry purification T. gondii tachyzoites and bradyzoites trypsin taurodeoxycholic acid |
| url | https://doi.org/10.1002/vms3.364 |
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