Generation of Humanized and Murinized NSG Mice

Female and male NSG (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ) mice from the Jackson Laboratories (Bar Harbor, ME, USA) or female Balb/c mice (BALB/cJRj, Janvier labs, Le Genest-Saint-Isle, France) were used for all experiments. Human CD34+ hematopoietic stem cells were isolated from human cord blood by magnetic separation (EasySep™ Human Cord Blood CD34 Positive Selection Kit II, STEMCELL technologies, Cologne, Germany) following the manufacturer’s protocol and purity controlled by flow cytometry. Humanized NSG (huNSG) mice were generated by engraftment of 100,000 hCD34+ cells (of a donor mix) at the age of 6 – 8 weeks, similar to the procedures described in earlier publications (15 (link)–18 (link)). Briefly, mice received whole-body irradiation with a sub-lethal dose of 2 Gy and hematopoietic stem cells were administered intravenously 2 hours later. For the generation of murinized NSG mice (muNSG), mice were treated as the huNSG mice, but received 100,000 bone marrow cells from a Balb/c donor instead of human CD34+ cells. The peripheral blood of huNSG mice was analyzed for the presence and frequency of murine CD45+, as well as human CD45+, CD3+ and CD20+ cells at week 18 post engraftment by flow cytometry.