Development of Onchocerca volvulus in humanized NSG mice and detection of parasite biomarkers in urine and serum

Patton, John B.; Bennuru, Sasisekhar; Eberhard, Mark L.; Hess, Jessica A.; Torigian, April; Lustigman, Sara; Nutman, Thomas B.; Abraham, David

The following human cell types were individually transferred into NSG mice: (1) human keratinocytes (HaCat) (ATCC, Manassas, VA), (2) bovine embryo skeletal muscle cells (BESM) In the initial experiments, mice were injected with 5×10 6 BESM, HaCaT, LEC, or HuSkMc cells subcutaneously weekly throughout the experiment. The frequency of injection was subsequently determined by in vivo imaging experiments. Genes encoding green fluorescent protein (GFP) and luciferase were inserted into HuSkMc cells using lentiviral vectors following the manufacturer's recommendations (Cell Biolabs, Inc, San Diego, CA). Cells expressing GFP were isolated using the GFP marker by fluorescence-activated cell sorting using a BD FACS Aria (BD Biosciences, Franklin Lakes, NJ). The isolated cells were grown in HuSkMc media as described above and 5×10 6 cells were injected subcutaneously into NSG mice. Mice were injected with VivoGlo (Promega, Madison, WI) and imaged following manufacturer's recommendations on an IVIS Lumina XR (Promega).