Figure 4.

CD38 knock-out CAR production process. Peripheral blood mononuclear cells were obtained by apheresis from a healthy volunteer. Cells were purified by CD3-negative selection using a CliniMACS system followed by a CD56-positive selection using MACS cell separation columns. After a short recuperation in IL2-supplemented media, the purified cells were subjected to CRISPR-Cas9 editing aimed to knock-out CD38. CD38 knock-out cells were then cultured for 14 days in an NAM-supplemented media in the presence of irradiated feeder cells and IL-15. On the day of harvest, an mRNA anti-CD38 CAR was introduced into cells by electroporation. Approximately 6 h post electroporation, the cells were cryopreserved.