This invention relates to the production of haemogenic endothelial cells (HECs). A population of induced pluripotent stem cells (iPSCs) is differentiated into mesoderm cells by culturing the IPSCs sequentially in first, second and third mesoderm induction media to induce differentiation into mesoderm cells. The mesoderm cells are then differentiated into HECs by culturing the mesoderm cells sequentially in first and second haemogenic endothelium (HE) induction media to induce differentiation into HECs. The HECs may be further differentiated into hematopoietic progenitor cells (HPCs) and progenitor T cells.

Provided is an isolated population of human pluripotent stem cells comprising at least 50% human pluripotent stem cells characterized by an OCT4+/TRA1-60?/TRA1-81?/SSEA1+/SSEA4? expression signature, and novel methods of generating and maintaining same in a pluripotent, undifferentiated state a suspension culture devoid of cell clumps. Also provided are novel culture media, cell cultures and methods for culturing pluripotent stem cells in a suspension culture or a two-dimensional culture system while maintaining the cells in a proliferative, pluripotent and undifferentiated state. The novel culture media comprise interleukin 11 (IL11) and Ciliay Neurotrophic Factor (CNTF); bFGF at a concentration of at least 50 ng/ml and an IL6RIL6 chimera; or an animal contaminant-free serum replacement and an IL6RIL6 chimera. Also provided are methods for generating lineage-specific cells from the pluripotent stem cells.

The present invention relates to methods, kits and compositions for expansion of embryonic hematopoietic stem cells and providing hematopoietic function to human patients in need thereof. In one aspect, it relates to kits and compositions comprising a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway. Also provided herein are methods for expanding embryonic hematopoietic stem cells using kits and compositions comprising a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway. The embryonic hematopoietic stem cells expanded using the disclosed kits, compositions and methods include cells derived from an embryo (e.g., aorta-gonad-mesonephros region of the embryo), embryonic stem cells, induced pluripotent stem cells, or reprogrammed cells of other types. The present invention also relates to administering the embryonic hematopoietic stem cells expanded using a combination of a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway to a patient for short-term and/or long-term in vivo repopulation benefits.

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.

Compositions and methods for manufacturing induced immune regulatory cells comprising induced myeloid suppressive cells including MDSCs (myeloid-derived suppressor cells), dendritic cells, macrophages, and subpopulations thereof are provided. Also provided are methods and compositions for further modifying and modulating the induced immune regulatory cells to achieve enhanced therapeutic potential in treating autoimmune disorders, hematological malignancies, solid tumors, viral infections, neurodegenerative disorders, inflammatory conditions, or GvHD.

Provided are defined serum-free culture media comprising a basal medium, serum replacement and an effective concentration of at least one differentiation inhibiting agent, wherein the defined culture medium is capable of maintaining mammalian livestock pluripotent stem cells in an undifferentiated state for at least 5 passages in culture, wherein the basal medium is selected suitable for maintaining pluripotent stem cells in an undifferentiated state, wherein the serum replacement comprises insulin and transferrin, and wherein the serum replacement is devoid of selenium. Also provided are methods of maintaining mammalian livestock pluripotent stem cells in an undifferentiated state, comprising culturing the mammalian livestock pluripotent stem cells in the defined culture medium.

A method for producing hematopoietic stem cells and/or hematopoietic progenitor cells from pluripotent stem cells is described. The method includes a step of culturing pluripotent stem cells in the presence of IGF2. A method is described for selecting an induced pluripotent stem cell(s) having high capacity to differentiate into hematopoietic stem cells and/or hematopoietic progenitor cells, or into blood cells, based on the expression level(s) of one or more genes such as TRIM58, CTSF, FAM19A5, and TCERG1L genes, or on the DNA methylation state(s) of the TRIM58, CSMD1, and/or FAM19A5 gene(s).

Described herein is a method for producing blood progenitor (hematopoietic progenitor cells) and T cell progenitor cells and to cells produced or obtainable by the process and the use of said cells, the method including: (a) optionally subjecting pluripotent stem cells under conditions that direct the cells to become mesoderm and subsequently hemogenic endothelial cells; and (b) directing hemogenic endothelial cells to differentiate into blood progenitor cells, preferably defined blood progenitor cells) using a media formulation designed to promote endothelial to hematopoietic transition (EHT) while being cultured on a surface functionalised with ligands designed to activate the Notch signaling pathway. In some aspects the ligands are Notch ligands, such as DLL4 and integrin ligands, such as integrin ?4?1 ligand or VCAM1.

An object of the present invention is to provide an ever-better method for producing sinusoidal endothelial cells. The present invention provides a method for producing sinusoidal endothelial cells from sinusoidal endothelial progenitor cells, the method comprising the step of culturing the sinusoidal endothelial progenitor cells in a medium containing one or more substances selected from the interleukin 6 (IL-6) family, for example, oncostatin M (OSM), interleukin 6 (IL-6), or interleukin 11 (IL-11).

Provided is an isolated population of human pluripotent stem cells comprising at least 50% human pluripotent stem cells characterized by an OCT4.sup.+/TRA1-60.sup.?/TRA1-81.sup.?/SSEA1.sup.+/SSEA4.sup.? expression signature, and novel methods of generating and maintaining same in a pluripotent, undifferentiated state a suspension culture devoid of cell clumps. Also provided are novel culture media, cell cultures and methods for culturing pluripotent stem cells in a suspension culture or a two-dimensional culture system while maintaining the cells in a proliferative, pluripotent and undifferentiated state. The novel culture media comprise interleukin 11 (IL11) and Ciliay Neurotrophic Factor (CNTF); bFGF at a concentration of at least 50 ng/ml and an IL6RIL6 chimera; or an animal contaminant-free serum replacement and an IL6RIL6 chimera. Also provided are methods for generating lineage-specific cells from the pluripotent stem cells.