P. falciparum parasites were cultured using human O+ erythrocytes in the presence of 10% pooled human A+ serum (Valley BioMedical, US) under standard conditions77 (link). The ItG-ICAM-1 parasite line78 (link), IT4var19 parasite line50 (link) and HB3var03 parasite line31 (link) were synchronized by alternating sorbitol treatment and gelatin selection. Mature trophozoite-stage IE (16–36 hrs post-invasion) or schizont-stage IE (>40 hrs and segmented) were enriched via magnet-column purification before being incubated with endothelial cell monolayers for 4–6 hrs or 12–20 hrs (overnight) at ratios ranging from 8–12 (106) to 80–120 (107) IE per endothelial cell. All parasite lines were regularly screened for mycoplasma negativity by PCR-based VenorGeM detection kit (Sigma, MP0025).
Venorgem detection kit
Manufactured by Merck Group
The VenorGeM detection kit is a laboratory equipment product designed for the detection and identification of various microorganisms. The kit provides a standardized and reliable method for the analysis of samples. It is intended for use by qualified laboratory professionals as part of their testing and research procedures.
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Lab products found in correlation
2 protocols using venorgem detection kit
1
Culturing and Synchronizing Malaria Parasites
2
Comprehensive Cell Culture Protocol
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B16-F10 melanoma cells (CRL-6045), 4T1 mammary carcinoma cells (CRL-2539), Lewis lung carcinoma (CRL-1642), and Vero kidney epithelial cells (CCL-81) were obtained from ATCC (Manassas, Virginia, USA). ID8 ovarian cancer cells (generated by Dr K Roby, University of Arkansas) were obtained from Dr Jean Marshall, Dalhousie University. Panc02 pancreatic cancer cells (generated by Dr T H Corbett, Wayne State University) were obtained from Dr John Bell, University of Ottawa. Cells were cultured at 37°C, 5% CO2, in Dulbecco’s Modified Eagle Medium (DMEM; VWR Life Science, Mississauga, ON, Canada) supplemented with 10% fetal bovine serum (FBS), 100 µg/mL streptomycin, and 100 units/mL penicillin (Fisher-HyClone, Ottawa, ON, Canada). Frozen stocks were generated, and aliquots were used in experiments at <8 passages. Mycoplasma testing was performed every 3–6 months using the VenorGem detection kit (Sigma-Aldrich).
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