Satellite cells and compositions and methods for producing the same
AssigneePresident and Fellows of Harvard College
InventorsLee L. Rubin, Feodor D. Price
Life of this patent
Estimating the term…
Abstract
Disclosed herein are methods for generating satellite cells and compositions including satellite cells.
Claims19
A method of generating a non-naturally occurring satellite cell comprising contacting a population of skeletal muscle organoids in three-dimensional spin culture with at least one differentiation medium comprising at least a cell culture medium and non-essential amino acids for at least 5 days to induce the dedifferentiation of at least one proliferative cell of the skeletal muscle organoids in the spin culture into a non-naturally occurring satellite cell, wherein the non-naturally occurring satellite cell expresses two or more genes selected from the group consisting of Nr2f1, Dmp1, Wisp2, Ecm1, Igfbp5, Itgb5, Sdc2, Bgn, and Mgp, and the two or more genes are not expressed in an endogenous satellite cell.
A method of generating a non-naturally occurring satellite cell comprising:
- contacting a population of cells comprising myoblasts with a myoblast medium to form an expanded myoblast population;
- contacting the expanded myoblast population in three-dimensional spin culture with a spin medium to form at least one skeletal muscle organoid comprising differentiated and proliferative cells;
- and
- contacting the at least one skeletal muscle organoid in three-dimensional spin culture with a differentiation medium comprising at least a cell culture medium and non-essential amino acids for a time sufficient to induce the dedifferentiation of at least one proliferative cell of the at least one skeletal muscle organoid into a non-naturally occurring satellite cell, wherein the non-naturally occurring satellite cell expresses two or more genes selected from the group consisting of Nr2f1, Dmp1, Wisp2, Ecm1, Igfbp5, Itgb5, Sdc2, Bgn, and Mgp, and the two or more genes are not expressed in an endogenous satellite cell.
A method of generating a non-naturally occurring satellite cell comprising:
- isolating a population of cells comprising myoblasts;
- contacting the population of cells comprising myoblasts with a myoblast medium comprising a cell growth medium, fetal bovine serum (FBS),fibroblast growth factor (FGF) and non-essential amino acids to form an expanded myoblast population;
- contacting the expanded myoblast population with a spin medium comprising a cell culture medium, FBS, FGF and non-essential amino acids for 10 to 30 days to form at least one skeletal muscle organoid comprising differentiated and proliferative cells;
- and
- contacting the at least one skeletal muscle organoid in spin culture with a differentiation medium comprising at least a cell culture medium and non-essential amino acids for at least 5 days to induce the dedifferentiation of at least one proliferative cell of the at least one skeletal muscle organoid in the spin culture into a non-naturally occurring satellite cell, wherein the non-naturally occurring satellite cell expresses two or more genes selected from the group consisting of Nr2f1, Dmp1, Wisp2, Ecm1, Igfbp5, Itgb5, Sdc2, Bgn, and Mgp, and the two or more genes are not expressed in an endogenous satellite cell.
Dependencies are inferred from the claim text.