NK cell compositions and preparations for immunotherapy and methods for their production

A method of preparing a natural killer (NK) cell composition or preparation, consisting of:

• selecting a plurality of umbilical cord blood or placental blood cells without immunological matching to each other;
• preparing enriched cluster of differentiation 34 positive (CD34+) hematopoietic stem and progenitor cells (HSPCs) that are depleted of red blood cells and T cells;
• culturing the CD34+ enriched HSPCs in an expansion culture medium comprising interleukin-3 (IL-3), interleukin-6 (IL-6), thrombopoietin (TPO), Fms-related tyrosine kinase 3 ligand (Flt-3 ligand (Flt-3L)), and stem cell factor (SCF) on a solid phase coated with a Notch ligand and recombinant human fibronectin or fragments thereof, and in the absence of exogenous feeder cells, for at least 7 days to produce expanded HSPCs, wherein the expanded HSPCs do not differentiate to produce more cluster of differentiation 56 positive (CD56+) cells than CD34+ cells during the expansion;
• and
• culturing the expanded HSPCs in a differentiation culture medium comprising effective amounts of interleukin-2 (IL-2) and interleukin-15 (IL-15), a non-animal sourced serum replacement, and in the absence of the exogenous feeder cells for at least 7 days to produce a NK cell composition or preparation comprising a population of NK cells of 50 to 80% CD56+ cells and 20 to 50% endogenous CD56 negative (CD56−) cells or 50 to 80% CD56+ cells and 15 to 50% endogenous CD56− cells;
• wherein the CD56+ cells express a high frequency of 60 to 100% of natural cytotoxicity receptor 3 (NKp30), natural cytotoxicity receptor 1 (NKp46), natural cytotoxicity receptor 2 (NKp44), immune inhibitory receptor natural killer group 2 member A (NKG2A), and granzyme B, a moderate to high frequency of 20 to 100% of perforin and lysosomal-associated membrane protein 1 (LAMP-1) or cluster of differentiation 107a (CD107a), a low to moderate frequency of 1 to 60% of cluster of differentiation 16 (CD16), and substantially no expression or less than 1% of killer-cell immunoglobulin-like receptors (KIRs);
• and wherein the CD56-cells express a moderate to high frequency of 20 to 100% of granzyme B, a high frequency of 60 to 100% of CD107a, and a low frequency of 1 to 20% of perforin.