Radial Glia and Astrocyte Differentiation From Human Pluripotent Stem Cells
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsIlyas Singec, Vukasin Jovanovic, Anton Simeonov
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Abstract
Methods for generating multipotent radial glia-like cells and astrocyte-like cells from human pluripotent stem cells are provided along with the related compositions.
Claims93
A method of producing, in culture, radial glia-like cells, the method comprising:
- (a) plating vertebrate pluripotent stem cells on a substrate-coated surface of a culture vessel at a density of 1,000-100,000 cells/cm2;
- (b) incubating the plated vertebrate pluripotent stem cells in a first culture medium;
- (c) replacing the first culture medium with a second culture medium comprising:
- (i) an effective amount or concentration of one or more inhibitors of BM P pathway,
- (ii) an effective amount or concentration of one or more activators of Notch pathway,
- (iii) one or more cytokines of interleukin-6 family;
- and
- (d) culturing the plated vertebrate pluripotent stem cells in the second culture medium;
- thereby producing radial glia-like cells.
A composition, comprising at least one cultured radial glia-like cell detectably expressing at least one marker, wherein the at least one marker is Brain Lipid Binding Protein (BLBP), CD133 (Prominin 1), abnormal spindle-like microcephaly-associated protein (ASPM), baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5 or Survivin), FAT Atypical Cadherin 1 (FAT1), Hes family bHLH transcription factor 5 (HES5), SRY-Box Transcription Factor 21 (SOX21), or PAX6 protein.
A cell culture, comprising at least one cultured radial glia-like cell detectably expressing at least one marker, wherein the at least one marker is Brain Lipid Binding Protein (BLBP), CD133 (Prominin 1), abnormal spindle-like microcephaly-associated protein (ASPM), baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5 or Survivin), FAT Atypical Cadherin 1 (FAT1), Hes family bHLH transcription factor 5 (HESS), SRY-Box Transcription Factor 21 (SOX21), or PAX6 protein.
A composition, comprising at least one cultured astrocyte-like cell exhibiting flat, star-shaped, and/or sphere morphology and detectably expressing at least one marker, wherein the at least one marker is S100 Calcium-Binding Protein B (S100B), Nuclear Factor 1 A-Type Protein (NFIA), Hepatic and Glial Cell Adhesion Molecule (HEPACAM), glial fibrillary acidic protein (GFAP), CD44 protein, or vimentin (VIM).
A cell culture, comprising at least one cultured astrocyte-like cell exhibiting flat, star-shaped, and/or sphere morphology and detectably expressing at least one marker, wherein the at least one marker is S100 Calcium-Binding Protein B (S100B), Nuclear Factor 1 A-Type Protein (NFIA), Hepatic and Glial Cell Adhesion Molecule (HEPACAM), glial fibrillary acidic protein (GFAP), CD44 protein, or vimentin (VIM).
Dependencies are inferred from the claim text.