Abstract: Disclosed are recombinant Cas9 proteins, methods of production, and methods of use for targeted DNA deletions, DNA insertions, or both in a eukaryotic genome. An assay system for evaluating the ability of the recombinant Cas9 proteins for targeted DNA deletions, DNA insertions, or both in a eukaryotic genome is also disclosed.

Abstract: Provided is a plate-shaped cartilage comprising a plurality of pluripotent stem cell-derived cartilaginous particles integrated with one another; and a method for producing the plate-shaped cartilage, comprising step 1: producing pluripotent stem cell-derived cartilaginous particles, and step 2: culturing the cartilaginous particles under culture conditions that allow adjacent cartilaginous particles to be in contact with one another, wherein the number of the cartilaginous particles is that amount required to form the plate-shaped cartilage.

Abstract: The present disclosure relates to a method of culturing quiescent hematopoietic stem cells. This method involves providing a culture medium and introducing, into the culture medium, quiescent hematopoietic stem cells to culture the stem cells and maintain quiescence of the stem cells. The culture medium comprises a vacuolar-H+ adenosine triphosphate ATPase (“v-ATPase”) inhibitor. Also disclosed are methods of treating a subject for a hematological disorder, methods of culturing leukemic stem cells, and methods of enhancing the hematopoietic reconstitution ability of a population of human hematopoietic stem cells.

Abstract: The invention relates to an aqueous solution for cell preservation of preferably mammalian cells, which can be used for cell preservation as a cryoprotectant or as a pharmaceutical product or excipient. Furthermore, the invention relates to a method for preserving cells using the aqueous solution for cell preservation, and to a method for defrosting cells which are frozen in the aqueous solution.

Abstract: Provided are therapeutic virus vectors, particularly, recombinant adeno-associated virus (rAAV) vectors, designed to contain an enhancer sequence that specifically restricts expression of an effector gene (e.g., an SCN1A-encoding polynucleotide, Gq-DREADD-encoding polynucleotide, or PSAM-encoding polynucleotide) contained in the vector to PV-expressing GABAergic interneuron or to neuron cell populations in the brain. The rAAV vectors, compositions and methods thereof are useful for treating subjects afflicted with neuropathologies, seizures, pharmacologically-intractable forms of epilepsy including Dravet syndrome (DS), a form of infantile epilepsy associated with severe seizures, cognitive impairment and premature death, as the cause of DS involves loss of function of a sodium channel encoded by the SCN1A gene.

Abstract: Disclosed herein are compositions comprising recombinant adeno-associated virus (rAAV), as well as recombinant baculovirus systems and methods of using the same for producing and purifying such compositions. Also disclosed herein are assays for testing the titer and potency of such compositions.

Abstract: The method for in vitro or ex vivo amplification of human adipose tissue stem cells includes: —extracting a stromal vascular fraction of a human adipose tissue including endothelial cells of the human adipose tissue vascular network and human adipose tissue stem cells, and an extracellular matrix of the human adipose tissue, the extracellular matrix including endothelial cells of the human adipose tissue vascular network, human adipose tissue stem cells and collagen; —mixing the stromal vascular fraction and the extracellular matrix; and —culturing the mixture obtained in the preceding step, in suspension, in a culture medium.

Abstract: The present invention relates to methods of efficiently generating pancreatic endoderm from human pluripotent stem (PS) cell derived human definitive endoderm. The present invention also relates to pancreatic endoderm cells obtained by the methods of the invention. Finally, the present invention relates to culture medium and composition comprising a RAR antagonist and uses of said RAR antagonist in the induction of pancreatic endoderm cells. The present invention provides a more homogenous and synchronised pancreatic cell population, with increased efficiency.

Abstract: The present invention relates generally to methods for genetic modification of hematopoietic cells. In particular, the invention relates to use of Prostaglandin E2 (PGE2), poloxamer, and protamine sulfate to enhance transduction by a recombinant retroviral vector. The compositions and methods of the present disclosure are particularly suitable for gene therapy applications, including the treatment of monogenic genetic diseases and disorders.

Abstract: IPEX (Immune dysregulation Polyendocrinopathy X linked) syndrome is a primary immunodeficiency caused by mutations in the gene encoding the transcription factor forkhead box P3 (FOXP3), which leads to the loss of function of thymus-derived CD4+CD25+ regulatory T (tTreg) cells. Preclinical and clinical studies suggest that T cell gene therapy approaches designed to selectively restore the repertoire of Treg cells by transfer of wild type FOXP3 gene is a promising potential cure for IPEX. However, there is still a need for a vector that can be used efficiently for the preparation of said Treg cells. The inventors thus compared 6 different lentiviral constructs according to 4 criteria (vector titers, level of transduction of human CD4+ T cells, level of expression of FOXP3 and ?LNGFR genes, degree of correlation between both expression) and selected one construct comprising a bidirectional PGK-EF1a promoter that showed remarkable efficiency.

Abstract: In certain embodiments, this disclosure provides novel methods of increasing the viable cell density of an N?1 large-scale bioreactor cell culture, comprising culturing a host cell expressing a recombinant poly peptide of interest in a non-perfusion-based culture system, and wherein the viable cell density is increased to at least 5×106 cells/mL. In certain embodiments, the disclosure provides novel methods for large-scale production of a recombinant polypeptide of interest, comprising: (1) culturing a host cell expressing a recombinant polypeptide of interest in an N?1 stage in a non-perfusion-based culture system, wherein the viable cell density is increased to at least 5×106 cells/mL; and (2) culturing N fed-batch production cells in an enriched media with high-seed density at least 1.5×106 cells/mL, wherein the N fed-batch production cells are inoculated from the N-1 stage in a non-perfusion-based culture system.

Abstract: Among the various aspects of the present disclosure is the provision of compositions and methods for platelet storage supplementation with an amino acid, such as taurine.

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: Disclosed herein are methods for treating or prophylactically treating age-related cognitive decline associated with dementia by administering recombinant viral vectors designed to deliver TERT and/or KL (Klotho) genes to the patient. The TERT and/or KL genes are packaged within one or more AAV viral vectors and administered to the CNS of the patient via intranasal and/or intrathecal injection. The therapy can provide improvements in cognitive function as evidenced by improved Folstein/MMSE scores over time.

Abstract: The present application relates to, inter alia, the identification, isolation and/or purification of cardiomyocytes in a sample. The method for isolating a cardiomyocyte population from a heterogeneous population of differentiated cells comprises: (a) contacting the sample with at least one agent that specifically binds to at least one cardiomyocyte surface marker selected from JAK2, DDR2, ACVRL1, CD200, SRPX, PRKACB, MST1R, P2RX1, TNFRSF10A, CHRND, KIAA0319, CD274, CCRL2, MBL2, ADORA3 and CD181; and (b) isolating the cells bound to the said agent. A preferred embodiment comprises contacting the sample with a first agent that specifically binds to a cell surface marker selected from JAK2, DDR2, ACVRL1, CD200, SRPX, PRKACB and MST1R to provide ventricular cardiomyocytes and a second agent that specifically binds to a cell surface marker selected from P2RX1, TNFRSF10A, CHRND, KIAA0319, CD274, CCRL2, MBL2, ADORA3 and CD181 to provide atrial cardiomyocytes.

Abstract: The invention relates to an activating composition comprising a cell, which may be any cell type used for cell therapy, wherein the cell is activated by a chemotherapy agent. Further, there is provided an activating composition comprising a supernatant of a composition comprising a cell, which may be any cell type used for cell therapy, wherein the cell is activated by a chemotherapy agent and wherein the supernatant is used as a therapy. The invention further provides methods for treating or preventing a disease or a condition comprising the use of the activated composition.

Abstract: Methods for the treatment of disorders associated with mitochondrial dysfunction, including rare, inborn errors of metabolism caused by genetic mutations; neurodegenerative disease; diabetes; and aging and age-associated decline. Generally, the methods include administering a therapeutically effective amount of one or more treatments that (i) induce hemoglobin “left-shifting” and/or (ii) induce anemia, as described herein, to a subject who is in need of, or who has been determined to be in need of, such treatment.

Abstract: Disclosed herein are compositions, constructs, cassettes, vectors, cells, nucleic acids, peptides, proteins, protocols and methods for reducing cholesterol and lipid buildup in mammalian subjects, via gene and/or cell therapeutic treatments. In many embodiments, the disclosed compositions, cells, constructs, cassettes, vectors, nucleic acids, peptides, proteins, protocols and methods may help to reduce lipid levels in mammals. In one embodiment, the disclosed compositions, cells, constructs, cassettes, vectors, nucleic acids, peptides, proteins, protocols and methods are useful in reducing lipid build-up, especially cholesterol, in liver cells.

Abstract: A method for producing pluripotent stem cells includes a step of performing suspension culture of pluripotent stem cells under a condition in which an amount, which is calculated by the following Equation (1), of WNT protein contained in a unit of a medium in contact with a unit area of a cell surface of a pluripotent stem cell is maintained at 2.9×102 ?g/mL·cm2 or less.

Abstract: The present invention generally relates to genetically modified swine wherein at least one allele of a SIGLEC1 gene has been inactivated and/or at least one allele of a CD163 gene has been inactivated. Genetically modified swine having both alleles of the SIGLEC1 gene and/or both alleles CD163 gene inactivated are resistant to porcine reproductive and respiratory syndrome virus (PRRSV). Methods for producing such transgenic swine are also provided.