The present invention relates to a method of culturing primitive-like macrophages from stem cells, a kit when used in the method thereof and uses of the primitive like macrophage for in-vitro disease models and for screening compounds for therapy. One embodied culture method comprises contacting and incubating embryonic stem cells or induced pluripotent stem cells with a serum-free culture media comprising a GSK3 inhibitor to differentiate stem cells into cells of the mesoderm lineage, followed by incubation with a culture media comprising Dickkopf-related protein 1 (DKK1) to differentiate the mesoderm into cells of hematopoietic lineage, maturing hematopoietic cells and incubating these cells with a culture media comprising M-CSF to drive differentiation into primitive-like macrophages. Another embodiment comprises incubating the stem cells with serum-free culture media comprising FGF2 and BMP4 to induce differentiation into cells of the mesoderm lineage, followed by incubating the cells with a culture media comprising FGF2, BMP4, Activin A and VEGF to differentiate the cells of the mesoderm lineage into cells of the hematopoietic cell lineage, maturing the cells of the hematopoietic cell lineage and lastly, incubating the matured hematopoietic cells with culture media comprising M-CSF to drive the differentiation of hematopoietic cells into primitive-like macrophages.

The present disclosure provides a method for preparing heterogeneous hematopoietic stem and progenitor cells using non-mobilized peripheral blood, which uses a capsule culture system to capture and proliferate rare hematopoietic stem and progenitor cells in non-mobilized peripheral blood, and prepares heterogeneous hematopoietic stem and progenitor cell clones. The present disclosure captures the rare heterogeneous stem cells in non-mobilized peripheral blood and morphologically verifies the presence of heterogeneous hematopoietic stem and progenitor cells in non-mobilized peripheral blood. The method of the present disclosure has the characteristics of hematopoietic reconstitution, drug development, transplantation and immunotherapy, gene editing of cell types, and the like. The method of the present disclosure provides a reliable cell source for patient-specific functional hematopoietic stem cells, and actively promotes the clinical application of non-mobilized hematopoietic stem and progenitor cells.

A method is provided for producing a plasmacytoid dendritic cells (pDCs), wherein hematopoietic stem and progenitor cells (HSPCs) are provided and incubated in a first medium comprising cytokines and growth factor whereby the HSPCs are differentiated into precursor-pDCs and then adding interferons (IFNs) to the first medium to obtain a second medium whereby said precursor-pDCs are differentiated into pDCs. Furthermore, a technique is provided for producing genetically modified pDCs, by initially genetically modifying HSPCs using transfection methods, including electroporation, to deliver sgRNA and Cas9 protein Moreover, a pharmaceutical formulation and a vaccine is provided which comprises pDC or genetically modified pDCs obtained according to that method.

The present invention provides a pluripotent stein cell comprising a co-expression vector in which Runx1 and Hoxa9 are of in tandem, and a T cell differentiated therefrom and application thereof. In the present invention, Pluripotent stein cells inducibly co-expressing exogenous Runx1 and Hoxa9 are successfully established by introducing an exogenous vector co-expressing Runx1 and Hoxa9 into pluripotent stein cells. The pluripotent stein cells are directionally differentiated into T-lineage progenitor cells and will be developed into T cells. The pluripotent stein cell-derived T cells obtained by the method of the present invention are not only functionally normal but also have no tumorigenic risk.

Multi-step methods for the in vitro production of enucleated red blood cells and the enucleated red blood cells thus prepared are provided. Such enucleated red blood cells may express fusion proteins comprising an antigen binding protein which allows the red blood cell to bind a toxin or an antigen of a pathogen. Also described herein are methods for neutralizing a toxin or pathogen in a subject by administering enucleated red blood cells that express any of the fusion proteins provided herein.

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.

A method of treating a respiratory viral infection in a mammal is provided. The method comprises administering to the mammal a therapeutically effective amount of alveolar-like macrophages, or an anti-viral factor produced by alveolar-like macrophages.

The present disclosure relates to artificial antigen presenting cells (aAPCs), in particular engineered erythroid cells and enucleated cells (e.g., enucleated erythroid cells and platelets), that are engineered to include exogenous-antigen presenting polypeptides comprising either HLA-E or HLA-G polypeptides, which may be used to activate or suppress certain immune cells.

The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.

The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.