Abstract: Embodiments of the disclosure concern methods and compositions related to optimization of selection of cord blood units to produce immune cells, such as NK cells, for adoptive cell therapy use. In specific embodiments, particular characteristics of the cord blood units and/or characteristics of cells derived therefrom are analyzed. When a threshold measurement for one or more characteristics is met for the cord blood unit(s) and/or characteristics of cells derived therefrom, the cord blood unit(s) are utilized as a source for production of immune cells. Specific characteristics for measurement include cord blood cell viability, total nuclear cell recovery, and nucleated red blood cell content, each prior to cryopreservation, and optionally cytotoxicity and/or expansion of immune cells subsequent to cryopreservation.

Abstract: Provided herein are methods of manufacturing clinical grade exosomes derived from mesenchymal stem cells (MSCs). Further provided are methods of loading the exosomes with therapeutic agents, such as siRNA. Also provided herein are methods of treating diseases by administering the clinical grade exosome.

Abstract: Embodiments of the disclosure include systems, methods, and compositions for producing megakaryocytes and platelets for recipient individuals in need thereof. The megakaryocytes and platelets are produced following co-culture of MSCs and CD34+ cells in media comprising stem cell factor, thrombopoietin, and IL-6, and wherein at least the CD34+ cells have a knock-in of HLA-E at the beta-2-microglobulin genomic locus, in specific embodiments. In some cases, ROCK inhibitors are utilized.

Abstract: Provided herein are methods of manufacturing clinical grade exosomes derived from mesenchymal stem cells (MSCs). Further provided are methods of loading the exosomes with therapeutic agents, such as siRNA. Also provided herein are methods of treating diseases by administering the clinical grade exosomes.

Abstract: Embodiments of the disclosure encompass systems, methods, and compositions for producing exosomes from primed mesenchymal stem cells that are expanded in the presence of IFN?, TNF?, IL-1?, and IL-17. The systems, methods, and compositions ay occur in an automated cell expansion system that allows for controllable parameters and from which cells and exosomes may be harvested at one or more times as part of a particular regimen. In specific embodiments, the exosomes may be provided to an individual in need thereof, including in some cases when the exosomes comprise one or more therapeutic agents.

Abstract: Embodiments of the disclosure includes methods of producing viral-specific therapy (VST) cells specific for the SARS-CoV-2 virus and uses of the cells. The methods may utilized peptide mixtures and stimulation of mononuclear cells using particular cytokine cocktails. The cells may also be genetically modified to lack expression of one or more endogenous genes, including one or more genes that renders the cells more effective and/or able to withstand deleterious conditions, such as the presence of glucocorticoids.

Abstract: Provided herein are chimeric antigen receptors (CARs) comprising a truncated EGFRvIII (Ev3) in the hinge region of the CAR and/or a humanized scFv. Further provided herein are immune cells expressing the CARs as well as methods of their use in the treatment of immune disorders.

Abstract: Embodiments of the disclosure encompass compositions comprising immune effector cells, such as natural killer (NK) cells, where the cells comprise one or more exogenously provided interleukins (IL), and wherein the cell optionally comprises one or more engineered receptors. In specific embodiments, the IL is not IL-15, and is IL-12, IL-21, or both. The NK cells may be utilized for treatment of cancer of any kind, including at least glioblastoma.

Abstract: Embodiments of the disclosure encompass methods and compositions for producing engineered mesenchymal stem/stromal cells (MSCs). The disclosure concerns large-scale processes for producing MSCs that are engineered to have disruption of expression of one or more genes using CRISPR and also express at least one heterologous antigen receptor. Specific embodiments include particular parameters for the process.

Abstract: Embodiments of the disclosure provide methods and compositions that facilitate cancer treatment including at least because they concern therapies that circumvent the tumor microenvironment. In specific embodiments, compositions are utilized for therapy that utilize NK cells that are protected from the direct inhibition of their activity (using TGF-beta inhibitors) and/or that are indirectly protected from TGF-beta (using integrin inhibitors). In specific embodiments, the NK cells have deficient expression and/or activity for TGF-beta Receptor 2 and/or glucocorticoid receptor.

Abstract: Provided herein are methods for expanding NK cells expressing chimeric antigen receptors and/or T cell receptors. Further provided are methods for treating diseases by administering the CAR NK cells.

Abstract: Provided herein are cryopreservation compositions and methods for cells of any kind, including for cells for adoptive cell therapy that are off-the-shelf cells. The cells for cryopreservation may be expanding NK cells expressing chimeric antigen receptors. In specific cases, the cryopreservation media comprises a cryoprotectant, such as DMSO, glycerol or hydroxyethol starch; serum or a non-serum alternative, such as platelet lysate; and one or more cytokines that are either natural, modified, synthetic, or recombinant.

Abstract: Provided herein are methods for expanding populations of regulatory B cells comprising engineering a population of B cells to express CD40 ligand. Also provided herein are methods of treating immune disorders with the regulatory B cells.

Abstract: Provided herein are methods for pre-activating and expanding an isolated population of NK cells. Further provided herein are methods for the treatment of cancer by administering the pre-activated and expanded NK cells.

Abstract: Provided herein are ex vivo methods for the expansion of cord blood-derived natural killer cells and methods of their use. Examples of embodiments include stimulating mononuclear cells from cord blood in the presence of antigen presenting cells (APCs) and IL-2 and re-stimulating the cells with APCs to produce expanded NK cells. In specific embodiments, the method does not utilize human leukocyte antigen (HLA) matching.

Abstract: Provided herein is a modular polycistronic vector system, such as for the genetic reprogramming of cells to express one or more antigen receptors. The modular characteristic of the system allows for exchange of one or more cistrons in addition to one or more components within particular cistrons. In specific embodiments, the modularity of the system allows exchange of components of a chimeric antigen receptor, such as exchange of costimulatory domains, scFvs, hinges, signaling domains, and so forth.

Abstract: Provided herein are methods for producing immune cells with disruption of multiple genes. Further provided are methods for inserting a chimeric antigen receptor at a gene locus of an immune cell.

Abstract: Embodiments of the disclosure encompass particular TNF-alpha mutants that are nonsecretable and membrane bound, thereby providing a target for inhibition in cells that express the mutants. In specific embodiments, the TNF-alpha mutants are utilized as a suicide gene in cells employed for adoptive cell therapy for an individual, wherein at a desired time the individual is provided one or more anti-TNF-alpha antibodies that bind the membrane bound TNF-alpha and elicit complement-dependent cytotoxicity for the cells. The TNF-alpha mutant can also be used as a way of tracking the transduced cells in vivo.

Abstract: Provided herein are methods for expanding populations of mesenchymal stromal cells (MSCs) comprising treating a population of MSCs derived from cord tissue with a pre-activation cytokine cocktail.

Abstract: The present invention concerns methods of treating a disease such as leukemia in a subject by administering natural killer (NK) cells. In particular aspects, HLA-C1-licensed KIR2DL2/3 and KIR2DS2 NK cells are administered to a subject with an HLA-C genotype either homozygous or heterozygous for the C1 allele, or HLA-C2 licensed cells are administered to a subject with an HLA-C genotype homozygous for the C2 allele. In further aspects, the NK cells are genetically modified to express a chimeric antigen receptor and interleukin 15.