We previously demonstrated manufacturing proof-of-concept, safety, and preliminary evidence of efficacy for ex vivoexpanded autologous and haploidentical NK cells in neuroblastoma, pediatric brain tumors, and myeloid leukemias. Despite initial success, the impact of these cellular therapies requiring bespoke manufacturing is limited by cost, manufacturing logistics, patient treatment delays, production failures, and donor availability. To resolve these issues, we worked with the National Marrow Donor Program to establish ideal NK cell-specific donor requirements and a screening, testing, and collection program with the aim of establishing a starting material bank for large-scale manufacturing of therapeutic NK cells. Our screening process evolved over time to settle on an initial screen based on HLA genotype, donor contact for interest, secondary screening for KIR genotype and infectious disease testing, tertiary screening for NK cell expansion and NKG2C expression, and a final screen to meet allogeneic donor requirements prior to collection of mononuclear cells by apheresis (MNC(A)). After collection, the MNC(A) is CD3-depleted, aliquoted, and cryopreserved. Manufacturing was optimized such that each aliquot is sufficient to generate up to 100x10⁹ NK cells after >5,000-fold expansion in 2 weeks, which are then aliquoted and cryopreserved, pass release testing, and may then be distributed for thawing and infusion at the bedside. Updates to donor screening, collection, and/or release testing have been implemented to meet new FDA guidance, accommodate non-US requirements (including Canada, Australia, Israel, and EU), or to incorporate novel assays for genomic integrity of gene-edited cells. We demonstrate a true off-the-shelf NK cell therapy program that has manufactured, distributed, and infused over 200 infusions of NK cells in over a dozen clinical trials at more than a dozen institutions under single-site and multi-site cooperative group studies, with sufficient material to deliver >5,000 doses at 10⁸/kg, and of broad HLA types to respond to alloreactivity.