Culturing and Propagating Type III Strains

Two type III strains were used in this study: 1) strain VEG; and 2) a CEP strain which had been engineered to contain a stably integrated GFP cassette [74 (link)]. Strains were propagated by passage in monolayers of primary human foreskin fibroblast cells (ATCC SCRC-1041), in Dulbecco’s modified Eagle’s medium (Wisent 319-015-CL) supplemented with 10% cosmic calf serum (HyClone SH30087.04), 20% M199 (Wisent 316-010-CL) and either penicillin/streptomycin (Wisent 450-201-EL) or, 10 U/mL penicillin and 10 μg/ml streptomycin (Gibco, Thermo Fisher Scientific Inc., Grand Island, NY), as previously described [75 (link)]. Genomic material was extracted, and PCR tested to ensure the lines were free from Mycoplasma infection. Tachyzoite-stage parasites were collected in 14 ml phosphate buffered saline (PBS) for injection into mice.

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Cromar G.L., Epp J.R., Popovic A., Gu Y., Ha V., Walters B.J., St. Pierre J., Xiong X., Howland J.G., Josselyn S.A., Frankland P.W, & Parkinson J. (2022). Toxoplasma infection in male mice alters dopamine-sensitive behaviors and host gene expression patterns associated with neuropsychiatric disease. PLoS Neglected Tropical Diseases, 16(7), e0010600.

Publication 2022

SCRC-1041 penicillin/streptomycin penicillin streptomycin

Cells fibroblast Cosmic Eagle Foreskin Genomic Human Mice Mycoplasma infection Parasites Penicillin Phosphate Saline Serum Strains Streptomycin

Corresponding Organization : University of Saskatchewan

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Variable analysis

independent variables

Type III strain used: 1) strain VEG; and 2) a CEP strain which had been engineered to contain a stably integrated GFP cassette

dependent variables

Not explicitly mentioned

control variables

Monolayers of primary human foreskin fibroblast cells (ATCC SCRC-1041) used for propagating the strains
Culture medium: Dulbecco's modified Eagle's medium (Wisent 319-015-CL) supplemented with 10% cosmic calf serum (HyClone SH30087.04), 20% M199 (Wisent 316-010-CL) and either penicillin/streptomycin (Wisent 450-201-EL) or, 10 U/mL penicillin and 10 μg/ml streptomycin (Gibco, Thermo Fisher Scientific Inc., Grand Island, NY)
Genomic material extracted and PCR tested to ensure the lines were free from Mycoplasma infection

controls

Positive control: Not mentioned
Negative control: Not mentioned

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