期刊
VETERINARY MEDICINE AND SCIENCE
卷 7, 期 2, 页码 357-361出版社
WILEY
DOI: 10.1002/vms3.364
关键词
flow cytometry; purification; T. gondii tachyzoites and bradyzoites; trypsin; taurodeoxycholic acid
资金
- Foshan University, Foshan city,Guangdong province
- Guangdong Science and Technology Plan Project [1244 0600 4560 7389XC]
- School of Life Science and Engineering fund [KLPREAD201801-02]
This study presents a simple and cost-effective method for large-scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites, which can be successfully reinfect HFF cells and propagate in mice. The new method allows for rapid preparation of sufficient quantity of pure parasites for downstream procedures.
This study describes a simple method for the large-scale isolation of pureToxoplasma gondiitachyzoites and bradyzoites.T. gondiitachyzoites 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 pureT. gondiitachyzoites 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.
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