Methods and compositions for generating oligodendrocyte progenitor cells
AssigneeTrailhead Biosystems Inc.
InventorNooshin Amini
Life of this patent
Estimating the term…
Abstract
Methods for generating pre-oligodendrocyte progenitor cells (pre-OPCs), oligodendrocyte progenitor cells (OPCs) and pre-myelinating oligodendrocytes (preOLs) from human pluripotent stem cells are provided using chemically-defined culture media that allow for generation of pre-OPCs in as little as three days, SOX10+ OLIG2+ NKX2-2+ OPCs in as little as twelve days and CD9+A2B5+O4+CNPase+ preOLs in as little as eighteen days. Two alternative culture protocols for generating OPCs are provided. Culture media, isolated cell populations and kits are also provided.
Claims26
A method of generating CD9+A2B5+O4+CNPase+ pre-myelinating oligodendrocytes (preOLs) comprising culturing SOX10+ OLIG2+ NKX2-2+ oligodendrocyte progenitor cells (OPCs) OPCs in a culture medium media comprising an insulin-like growth factor 1 receptor (IGF1R) pathway agonist, a tropomyosin-related kinase C (TrkC) pathway agonist, a platelet-derived growth factor receptor (PDGFR) pathway agonist, a thyroid hormone receptor agonist, and an insulin receptor agonist such that CD9+A2B5+O4+CNPase+ preOLs are generated.
A method of generating CD9+A2B5+O4+CNPase+ preOLs comprising:
- (a) culturing human OLIG2+ pre-oligodendrocyte progenitor cells (pre-OPCs) in a culture medium comprising a fibroblast growth factor receptor (FGFR) pathway agonist, a mammalian target of rapamycin (mTOR) pathway antagonist, a sonic hedgehog (SHH) pathway agonist, an Akt pathway antagonist, and an Akt pathway agonist on days 0-3 to obtain a population of cells;
- (b) culturing the population of cells from step (a) in a culture medium comprising an FGFR pathway agonist, a PDGFR pathway agonist, an Akt pathway antagonist, and a retinoic acid (RA) pathway agonist on days 3-9, such that SOX10+ OLIG2+ NKX2-2+ OPCs are generated;
- and
- (c) culturing the population of cells from step (b) in a culture medium comprising an IGF1R pathway agonist, a TrkC pathway agonist, a PDGFR pathway agonist, a thyroid hormone receptor agonist, and an insulin receptor agonist on days 9-15 such that CD9+A2B5+O4+CNPase+ preOLs are generated.
A method of generating human CD9+A2B5+O4+CNPase+ preOLs comprising:
- (a) culturing human OLIG2+ pre-OPCs in a culture medium comprising an FGFR pathway agonist, an mTOR pathway antagonist, an SHH pathway agonist, and a WNT pathway agonist on days 0-6 to obtain a population of cells;
- and
- (b) culturing the population of cells from step (a) in a culture medium comprising an FGFR pathway agonist, an IGF-1 pathway agonist, and an RA pathway agonist on days 6-9, such that SOX10+ OLIG2+ NKX2-2+ OPCs are generated;
- and
- (c) culturing the population of cells from step (b) in a culture medium comprising an IGF1R pathway agonist, a TrkC pathway agonist, a PDGFR pathway agonist, a thyroid hormone receptor agonist, and an insulin receptor agonist on days 9-15 such that CD9+A2B5+O4+CNPase+ preOLs are generated.
Dependencies are inferred from the claim text.