METHOD FOR DIFFERENTIATING PLURIPOTENT STEM CELLS INTO NATURAL KILLER CELLS

Claims

The present invention describes a method for substantially expanding the natural killer (NK) cell population using modified cell culture conditions. Employing a targeted induction cocktail, which includes a WNT signaling pathway activator and bone morphogenetic protein (BMP), this technique facilitates the differentiation of pluripotent stem cells (PSCs) into NK cells. The core innovation lies in optimizing the culture conditions, which significantly increases NK cell proliferation. This strategy accelerates the production of NK cells for therapeutic uses, emphasizing the expansion phase within the culture environment.

1. A method for producing natural killer (NK) cells comprising: a) contacting a pluripotent stem cell (PSC) culture with an activator of the WNT signaling pathway and / or a bone morphogenetic protein (BMP), wherein the PSCs are cultured for approximately 1-7 days; b) contacting the PSC culture with vascular endothelial growth factor (VEGF) for approximately 1-7 days following step a), thereby generating a population of CD34+ precursor cells enriched to at least approximately 80% CD34+;c) incubating the CD34+ precursor cell population for approximately 7-28 days in a culture medium lacking any serum-derived agent while optionally supplementing the medium with one or more of interleukin-7 (IL-7), optionally IL-15 NK-activating cytokine, stem cell factor (SCF), and FMS-like tyrosine kinase ligand 3 (FLT3L), thereby creating a CD34+ / CD45+ cell suspension population; and d) incubating the CD34+ / CD45+ cell suspension population in a culture medium containing at least one serum-derived agent and IL-7, optionally IL-15 NK-activating cytokine, SCF, and FLT3L for approximately 7-35 days, thereby producing NK cells.