Ipsc-derived immune cells in prophylaxis and treatment of age-associated and neurodegenerative diseases

A method for generating mononuclear phagocytes comprising:

• culturing induced pluripotent stem cells in a cell culture medium under conditions that induce myeloid differentiation, leading to the generation of mononuclear phagocytes, wherein the culturing comprises contacting the induced pluripotent stem cells with a first composition comprising bone morphogenetic protein (BMP-4),
• contacting the cell culture medium with a second composition after culturing of the induced pluripotent stem cells in the presence of the first composition, said second composition comprising one or more factors selected from the group consisting of bFGF, vascular endothelial growth factor (VEGF), and stem cell factor (SCF), and a combination thereof,
• contacting the cell culture medium with a third composition after culturing in the presence of the second composition, said third composition comprising one or more factors selected from the group consisting of SCF, interleukin 3 (IL-3), thrombopoietin (TPO), macrophage colony-stimulating factor (M-CSF), and Fms-like tyrosine kinase 3 ligand (FLT3 ligand), and a combination thereof, and
• contacting the cell culture medium with a fourth composition after culturing in the presence of the third composition, said fourth composition comprising one or more factors selected from the group consisting of M-CSF, GM-CSF, and FLT3 ligand, and a combination thereof, thereby generating the mononuclear phagocytes,
• wherein the culturing does not include contacting the generated mononuclear phagocytes in a microglial differentiation medium comprising interleukin 34 (IL-34) or comprising IL-34 and granulocyte-macrophage colony-stimulating factor (GM-CSF), and the culturing does not include contacting the generated mononuclear phagocytes in dendritic cell differentiation medium comprising interleukin 4 (IL-4) or comprising IL-4 and GM-CSF.

Mononuclear phagocytes generated using a method comprising:

• (1) culturing pluripotent stem cells in an adherent culture in a first cell culture medium comprising bone morphogenetic protein 4 (BMP-4);
• (2) culturing the cells obtained by step (1) in an adherent culture in a second cell culture medium comprising basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and stem cell factor (SCF);
• (3) culturing the cells obtained by step (2) in an adherent culture in a third cell culture medium comprising SCF, interleukin 3 (IL-3), thrombopoietin, macrophage colony-stimulating factor (M-CSF), and FLT3 ligand;
• and
• (4) culturing the cells obtained by step (3) in a suspension culture in a fourth cell culture medium comprising M-CSF, granulocyte-macrophage colony-stimulating factor (GM-CSF), and FLT3 ligand, wherein macrophages and/or monocytes are generated;
• wherein the method does not include contacting the generated mononuclear phagocytes in a microglial differentiation medium comprising interleukin 34 (IL-34) or comprising IL-34 and GM-CSF, and the culturing does not include contacting the generated mononuclear phagocytes in dendritic cell differentiation medium comprising interleukin 4 (IL-4) or comprising IL-4 and GM-CSF;
• and
• wherein the mononuclear phagocytes are positive for CD11b, CD14, CD16, CD64, CD11c, CD71, or a combination thereof.

A method for generating mononuclear phagocytes and treating a subject with a neurodegenerative disorder or a subject in need of cognitive function improvement, comprising:

• culturing induced pluripotent stem cells in a cell culture medium under conditions that induce myeloid differentiation, leading to the generation of mononuclear phagocytes, wherein the culturing comprises contacting the induced pluripotent stem cells with a first composition comprising bone morphogenetic protein (BMP-4),
• contacting the cell culture medium with a second composition after culturing of the induced pluripotent stem cells in the presence of the first composition, said second composition comprising one or more factors selected from the group consisting of bFGF, vascular endothelial growth factor (VEGF), and stem cell factor (SCF), and a combination thereof,
• contacting the cell culture medium with a third composition after culturing in the presence of the second composition, said third composition comprising one or more factors selected from the group consisting of SCF, interleukin 3 (IL-3), thrombopoietin (TPO), macrophage colony-stimulating factor (M-CSF), and Fms-like tyrosine kinase 3 ligand (FLT3 ligand), and a combination thereof, and
• contacting the cell culture medium with a fourth composition after culturing in the presence of the third composition, said fourth composition comprising one or more factors selected from the group consisting of M-CSF, GM-CSF, and FLT3 ligand, and a combination thereof, thereby generating the mononuclear phagocytes,
• wherein the culturing does not include contacting the generated mononuclear phagocytes in a microglial differentiation medium comprising interleukin 34 (IL-34) or comprising IL-34 and granulocyte-macrophage colony-stimulating factor (GM-CSF), and the culturing does not include contacting the generated mononuclear phagocytes in dendritic cell differentiation medium comprising interleukin 4 (IL-4) or comprising IL-4 and GM-CSF;
• and
• administering to the subject a therapeutically effective amount of a composition comprising the generated mononuclear phagocytes, thereby treating the subject with the neurodegenerative disorder or improving the cognitive function in the subject.