Methods and compositions for reprogramming cells
AssigneeThe Regents of the University of Colorado, a body corporate
InventorsIgor Kogut, Dennis R. Roop, Ganna Bilousova
Life of this patent
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Abstract
The present disclosure relates to methods and compositions for reprogramming cells to a pluripotent state. In particular, it relates to an integration- and feeder cell-free method for reprogramming primary human fibroblast cells to induced pluripotent stem cells (iPSCs).
Claims20
A reprogramming composition for reprogramming primary fibroblast cells to a pluripotent state, the composition comprising:
- primary fibroblast cells at a density of less than about 1000 cells/cm2 or less in the absence of feeder cells and in the absence of treated serum albumin;
- a complexation buffer of pH 7.6 to less than pH 8.6;
- and
- a reprogramming RNA cocktail, wherein the reprogramming RNA cocktail comprises mRNA encoding Sox2, Klf4, and at least one of Myo-D-Oct4 (M3O) or Oct4.
A composition for reprogramming a single primary human fibroblast
- cell to a pluripotent state,
- the composition comprising:
- a single cell in the absence of feeder cells and in the absence of treated serum albumin:
- a complexation buffer;
- and
- a reprogramming RNA cocktail comprising:
- 100 to 1,500 ng/10 cm2 reprogramming RNA cocktail and 5 to 40 pmoles/10 cm2 reprogramming miRNA, wherein pH of the complexation buffer is 7.8 to 8.4 and wherein the reprogramming RNA cocktail comprises mRNA encoding Sox2, Klf4, and at least one of Myo-D-Oct4 (M3O) or Oct4.
A composition for reprogramming primary fibroblast cells to a pluripotent state, the composition comprising primary fibroblast cells at a density of about 1000 cells/cm2 or less in the absence of feeder cells and in the absence of treated serum albumin;
- a complexation buffer of pH 7.6 to less than pH 8.6;
- and
- a reprogramming RNA cocktail, wherein the reprogramming RNA cocktail comprises mRNA encoding Sox2, Klf4, c-Myc, Lin28A, and at least one of Myo-D-Oct4 (M3O) or Oct4.
Dependencies are inferred from the claim text.