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US 12,097,223 B2Granted
Mesenchymal stromal cells and uses related thereto
AssigneeAstellas Institute for Regenerative Medicine
InventorsErin Anne Kimbrel, Robert P. Lanza, Jianlin Chu, Nicholas Arthur Kouris
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Abstract
The present invention generally relates to novel preparations of mesenchymal stromal cells (MSCs) derived from hemangioblasts, methods for obtaining such MSCs, and methods of treating a pathology using such MSCs. The methods of the present invention produce substantial numbers of MSCs having a potency-retaining youthful phenotype, which are useful in the treatment of pathologies.
Claims9
A method for generating human mesenchymal stromal cells, the method comprising
- (a) differentiating human pluripotent stem cells in vitro in a first medium comprising 20-100 ng/mL vascular endothelial growth factor (VEGF) and 15-100 ng/mL bone morphogenic protein 4 (BMP-4) to generate cell clusters,
- (b) culturing the cell clusters in a second medium comprising 20-100 ng/mL VEGF, 15-100 ng/mL BMP-4, and 20-25 ng/mL basic fibroblast growth factor (bFGF),
- (c) dissociating the cell clusters to obtain single cells,
- (d) culturing the single cells in a serum free, methylcellulose-based medium to generate hemangioblasts, and
- (e) differentiating the hemangioblasts in vitro to generate human mesenchymal stromal cells, wherein said hemangioblasts are cultured in the presence of transforming growth factor beta (TGF-beta), epidermal growth factor (EGF), insulin-like growth factor 1, basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), or a combination thereof.
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