Method of forming myeloid precursor cells from human embryonic stem cells
AssigneeWisconsin Alumni Research Foundation
InventorsIgor I. Slukvin, James A. Thomson, Maksym A. Vodyanyk, Maryna E. Gumenyuk
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Abstract
This invention relates to the culture of dendritic cells from human embryonic stem (ES) cells. Human ES cells are first cultured into hematopoietic cells by co-culture with stromal cells. The cells now differentiated into the hematopoietic lineage are then cultured with GM-CSF to create a culture of myeloid precursor cells. Culture of the myeloid precursor cells with the cytokines GM-CSF and IL-4 causes functional dendritic cells to be generated. The dendritic cells have a unique phenotype, as indicated by their combination of cell surface markers.
Claims3
A culture of myeloid precursor cells in which a majority of the cells have a phenotype of myeloid precursors and in which at least 90% of the cells are CD45+, CD4+, CD123low, negative for HLA-DR and include subpopulations of cells expressing MPO, M-CSFR, CD11b, CD11c, CD15 and CD16.
A method of obtaining myeloid precursor cells comprising the steps of:
- (a) co-culturing human embryonic stem cells with stromal cells that do not express macrophage colony-stimulating factor, wherein the stem cells are induced to differentiate into multipotent lympho-hematopoietic progenitor cells and wherein the culture is free of exogenously added cytokines;
- (b) culturing the multipotent lympho-hematopoietic progenitor cells of step (a) with granulocyte/macrophage colony stimulating factor (GM-CSF) under non-adherent conditions to produce and expand myeloid precursor cells, wherein at least 90% of the resultant cells are myeloid precursor cells expressing CD45+, CD4+, CD123low, negative for HLA-DR and include subpopulations of cells expressing MPO, M-CSFR, CD11b, CD11c, CD15 and CD16.
Dependencies are inferred from the claim text.