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El-Ashram S, Zhang Y, Ji Y, Salama D, Mei K, Zhili L, Shujian H, Zhang H, Aboelhadid SM, Alajmi RA, Metwally DM, El-Khadragy MF, Hargis BM, Tellez-Isaias G, Cenci-Goga BT, Karama M, Marufu MC, Abouhajer F, Ali Abdelhafez Hamady G, El Wakil A, Al Nasr I, Suo X.
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Vet Med Sci. 2021 Mar;7(2):357-361. doi: 10.1002/vms3.364. Epub 2020 Sep 26.
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. CI – © 2020 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.
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Link/DOI: 10.1002/vms3.364