A method for producing CD4/CD8 double-positive T cells, comprising the steps of: (1) culturing pluripotent stem cells in a medium to induce hematopoietic progenitor cells; and (2) culturing the hematopoietic progenitor cells obtained in the step (1) in a medium containing a p38 inhibitor and/or SDF-1 to induce CD4/CD8 double-positive T cells.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor and/or survival factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes a survival factor, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor and/or a different survival factor.

Provided is a human-derived sensory neuron induction culture system. A combination of a small molecule inhibitor LY2157299 and a growth factor is added into a serum-free basal medium. Compared with an induction method involving serum, not only is the efficiency of inducing pluripotent stem cells into sensory neurons greatly improved, but the expression of a variety of ion channel proteins is also significantly improved, thereby achieving successful induction of multiple induced pluripotent stem cells from different sources into sensory neurons.

The present invention relates to a stable method for introducing at least one inducible cassette into a cell, and permitting controllable transcription from within that inducible cassette. The method may be used for any cell type, from any eukaryotic organism, but has a particular application in the introduction of inducible cassettes into pluripotent stem cells, such as animal or human pluripotent stem cells (hPSCs). The inducible cassette is controllably inserted in such a way to ensure that the genetic material it contains is not silenced or subject to negative influences from the insertion site, and transcription of the genetic material is controlled.

Provided herein are methods and compositions for engineering antigen-presenting cells (APCs), such as dendritic cells (DCs), macrophages, and B-Cells, that are modified to express viral antigens. With this approach, large cell banks can be created that can be rapidly modified and deployed to fight various types of infectious diseases such as those associated with coronaviruses and other viral infections.

Maturation signals provided via cyclic adenosine monophosphate (cAMP)/Exchange proteins activated by cAMP (Epac) signaling during in vitro generation of blood cells from reprogrammed cells or pluripotent stem cells achieve superior function of hematopoietic cells differentiated from stem cells. The cAMP/Epac signaling enables an increased efficiency of production of precursor to blood and to blood cells. These generated blood cells can be utilized for therapeutics, treatments, disease prevention, drug discovery, personalized medicine, regenerative medicine, cell and tissue generation, universal donor banks and related methods and compositions.

The present specification provides: a method of isolation of a pure culture of vascular endothelial cells, the method capable of isolating homogeneous endothelial cells adhered to a matrix for a specific time in a cell line of an endothelial cell lineage differentiated from human pluripotent stem cells; a medium for maintaining characteristics of vascular endothelial cells, comprising high-purity vascular endothelial cells isolated through the method, 4 ng/ml to 6 ng/ml of FGF2, 5 ng/ml to 10 ng/ml of EGF, 10 ng/ml to 30 ng/ml of VEGF-A, 20 ng/ml to 50 ng/ml of ascorbic acid, and DMEM/F-12 as active ingredients; and a culture method comprising same.

Embodiments of the disclosure concern methods and compositions related to cancer treatment for an individual utilizing recombinant fibroblast cells that comprise one or more activities that are endothelial cell-like. The cells are delivered to a tumor microenvironment following which their death results in destabilization of the tumor vasculature. In particular embodiments, the fibroblast cells recombinantly express one or more of ETV2, FOXC2, and FLI1.

The disclosure features methods and compositions for differentiating stem cells into hematopoietic stem and progenitor cells (HSPC) and/or Natural Killer (NK) cells. The methods and compositions described herein are used to differentiate stem or progenitor cells having at least one gene-edit that is maintained in the differentiated cell. Also provided are differentiated cells produced using the methods and compositions described herein for therapeutic applications.

The present invention provides in vitro methods of differentiating brain mural cells and methods of use, including use in blood brain barrier models. Suitable in vitro derived cell populations of brain mural cells are also provided.