Bioluminescent PDX model of B-ALL

We created PDX models that developed stable bioluminescence by infecting B-ALL cells #2 with a lentivirus expressing both GFP and luciferase [67 (link)]. Then, we transplanted these cells into mice that were used later for bioluminescence imaging. The lentivirus was produced in HEK293 cells after transduction with Lipofectamin 2000 (Thermo Fisher Scientific) of the pCCLc-MNDU3-Luciferase-PGK-EGFP-WPRE vector (Addgene, #89608), as well as PAX2 (Addgene, #12260) and pCMV-VSV-G (Addgene, #8454) plasmids. After 2 days, viral supernatants were recovered, and 6-well plates were incubated for 4 h with retronectin (Takara, Ozyme). Viral supernatants were then spinoculated for 30 min at 4000×g. Cells were cultured on these plates for three days in StemMACS media (Miltenyi Biotech). Lentiviral transduced cells (GFP⁺) were sorted on a FACSAriaIII cell sorter (BD Biosciences) and transplanted in NSG mice to generate bioluminescent PDX. Following isoflurane-induced anesthesia, animals were imaged 15 min after d-luciferin (Merck) injection, at 150 mg/kg body weight, using an IVIS Lumina III system coupled with Living Image acquisition and analysis software version 4.0 (Perkin Elmer).