The present invention relates to, inter alia, an engineered cell (e.g., iPSC, IPS-derived NK, or NK cell) comprising a disrupted B2M gene and an inserted polynucleotide encoding one or more of SERPINB9, a fusion of IL15 and IL15Rα, and/or HLA-E. The engineered cell can further comprise a disrupted CIITA gene and an inserted polynucleotide encoding a CAR, wherein the CAR can be an anti-BCMA CAR or an anti-CD30 CAR. The engineered cell may further comprise a disrupted ADAM17 gene, a disrupted FAS gene, a disrupted CISH gene, and/or a disrupted REGNASE-1 gene. Methods for producing the engineered cells are also provided, and therapeutic uses of the engineered cells are also described. Guide RNA sequences targeting described target sequences are also described.

In some embodiments, the present invention provides tissue grafts, such as vascularized bone grafts, and methods for preparing and using such tissue grafts. In some embodiments the tissue grafts are made using pluripotent stem cells, such as autologous pluripotent stem cells. In some embodiments, the tissue grafts are made by creating a digital model of a tissue portion to be replaced or repaired, such as a bone defect, partitioning the model into two or more model segments, and then producing tissue graft segments having a size and shape corresponding to that of the model segments. Such tissue graft segments may be assembled to form a tissue graft having a size and shape corresponding to that of the tissue portion to be replaced or repaired.

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.

A non-degradable device for use in controlled feeding of mammalian cell cultures including by way of example cultures of stem cells such as induced pluripotent stem cells (iPSCs). Methods of making and using the device are also disclosed.

A method to enhance homology directed recombination (HDR) efficiency, and a kit therefor, are provided.

Disclosed are T regulatory cells endogenously occurring or generated in vitro which possess regenerative activity. In one embodiment said T regulatory cells are created by culture with derivatives of activated/enhanced mesenchymal stem cells. Said derivatives include apoptotic bodies, conditioned media, exosomes, microvesicles, proteins and various metabolites. In one embodiment said mesenchymal stem cells are activated with cytokines such as interferon gamma. In other embodiments are said mesenchymal stem cells are activated by ligation of toll like receptors. Cells of the invention are useful for treatment of autoimmune, degenerative, and combination of autoimmune and degenerative disease. Other uses of said regenerative T cells include treatment of heart failure, liver failure, stroke, and ischemic diseases.

The invention relates to a method for the formation of renal tubules by embedding individual renal cells into a synthetic hydrogel, which is based on polyethylene glycol as a component, and the culturing of the cells until tubule structures are formed. The culturing can be continued until the obtained tubule structures correspond in terms of size, structure, morphology and functionality to adult human renal tubules or are at least similar thereto.

The invention provides culture platforms, cell media, and methods of differentiating pluriptent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, and NK cells.

The present invention relates to a composition for the differentiation of dental pulp stem cells into odontoblasts and an IgG or IgM type monoclonal antibody that specifically binds to the surface of odontoblasts differentiated from the stem cells. According to the present invention, BMP2 and BMP4 are optimally combined to significantly increase the differentiation efficiency of dental pulp stem cells into odontoblasts, to induce the mineralization of the matrix, and to improve the differentiation ability of odontoblasts into dentin. Further, the IgG or IgM monoclonal antibody that specifically binds to the surface of odontoblasts of the present invention is used to effectively isolate and purify odontoblasts, which can be useful for tissue regeneration and differentiation.

Provided herein are compositions and methods for generation of naive human pluripotent stem cells. The method comprises incubation of iPSCs under 5% O.sub.2 in a medium comprising 5% glucose, an MEK inhibitor, a GSK3β inhibitor, human leukemia inhibitory factor (LIF), human insulin and Torin 1. The method does not need any other inhibitors or transgene expression. The naive human pluripotent cells can be used to generate a large amount of mature human cells from all three germ layers in host non-human animals.