Embodiments herein relate to in vitro production methods of hematopoietic stem cell (HSC) and hematopoietic stem and progenitor cell (HSPC) that have long-term multilineage hematopoiesis potentials upon in vivo engraftment. The HSC and HSPCs are derived from pluripotent stem cells-derived hemogenic endothelia cells (HE) by non-integrative episomal vectors-based gene transfer.

The invention relates to a method of producing an iPSC-derived Kupffer Cell (IKC). The method may comprise providing a macrophage precursor (preMcp) derived from an induced pluripotent stem cell (iPSC). The macrophage precursor (preM-cp) may be cultured in the presence of a hepatic cue, such as a combination of primary human hepatocyte conditioned media and Advanced DMEM, thereby obtaining the iPSC-derived Kupffer Cell. The iPSC-derived Kupffer Cell may display a biological property of a primary Kupffer cell, such as a primary adult human KC (pKC). The biological activity comprises expression of a macrophage marker such as CD11, CD14, CD68, CD163, CD32, CLEC-4F, ID1 and ID3.

The present invention provides a cell population and a method of obtaining the same. The cell population of the present invention is obtained by culturing mononuclear cells derived from bone marrow, umbilical cord blood, or peripheral blood in a medium containing serum and four or less of factors selected from the group consisting of stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 ligand, thrombopoietin, and vascular endothelial growth factor.

Methods of generating definitive hematopoietic cells from source cells including at least one of: differentiating iPS cells, cells directly reprogrammed to pre-cursors of hematopoietic cells, cells directly reprogrammed to definitive hematopoietic cells, and adult or neonatal hematopoietic cells from bone marrow, cord blood, placenta, or mobilized peripheral blood, the method including using a metabolic regulator to activate a tricarboxylic acid cycle of the source cells. Other methods relate to generating primitive hematopoietic cells from source cells including at least one of: differentiating iPS cells, cells directly reprogrammed to pre-cursors of hematopoietic cells, cells directly reprogrammed to definitive hematopoietic cells, and adult or neonatal hematopoietic cells from bone marrow, cord blood, placenta, or mobilized peripheral blood, the method including using a metabolic regulator to inhibit a tricarboxylic acid cycle of the source cells. Some aspects relate to a metabolic regulator for activation of a tricarboxylic acid cycle of source cells for the production of definitive or primitive hematopoietic cells.

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.

Provided are methods for expanding stem cells and/or lineage committed progenitor cells, such as hematopoietic stems cells and/or lineage committed progenitor cells, at least in part, by using compounds that antagonize AhR. The compounds are represented by formulae:

##STR00001## wherein the letters and symbols a, b, c, d, e, f, g, Z, R.sup.1b, R.sup.2a and R.sup.2b have the meanings provided in the specification. Also provided are compositions comprising stem cells and/or lineage committed progenitor cells expanded by methods disclosed herein and methods for the treatment of diseases treatable by same.

A method of making an erythroid cell comprising elevated levels of a target protein or polypeptide, the method comprising: a) provision of an erythroid progenitor which is able to express the target protein or polypeptide; b) expression of the target protein or polypeptide; and c) maturation of the erythroid progenitor into the erythroid cell; wherein during maturation of the erythroid progenitor into the erythroid cell, the target protein or polypeptide is configured and/or inhibited such that ubiquitination of the target protein or polypeptide is hindered or prevented. Erythroid cells, pharmaceutical compositions and methods of use related thereto, and a method of screening for proteins or polypeptides degraded by ubiquitination during maturation of an erythroid progenitor are also provided.

The disclosure relates to stem cells and their therapeutic use in the treatment and/or prevention of pancreatic diseases or disorders. Provided herein are compositions comprising c-kit positive pancreatic stem cells and methods of preparing and using c-kit positive pancreatic stem cells for the treatment and/or prevention of pancreatic diseases or disorders.

A method for producing a professional antigen-presenting cell, including inducing expression of c-MYC, BMI1, and MDM2 in a myeloid cell (MC) to obtain a proliferative myeloid cell (pMC), and inducing expression of GM-CSF and/or M-CSF in the pMC to obtain a professional antigen-presenting cell (pAPC). The myeloid cell is a myeloid cell differentiated from a pluripotent stem cell.

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.