Veterinary medicine and science · 2020
A rapid and simple single-step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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El-Ashram Saeed, Zhang Yu, Ji Yongsheng, Salama Dina, Mei Kun, Zhili Li, Shujian Huang, Zhang Haoji, Aboelhadid Shawky M, Alajmi Reem A, Metwally Dina M, El-Khadragy Manal F, Hargis Billy M, Tellez-Isaias Guillermo, Cenci-Goga Beniamino T, Karama Musafiri, Marufu Munyaradzi C, Abouhajer Fathi, Ali Abdelhafez Hamady Gamal, El Wakil Abeer, Al Nasr Ibrahim, Suo Xun
Аннотация
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.