Methods and products for transfection
The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.
1. A method for reprogramming a cell to a less differentiated state, comprising:
(a) providing a non-pluripotent cell;
(b) culturing the cell in a medium containing ingredients that support reprogramming of the cell to a less differentiated state; and
(c) transfecting the cell with a synthetic RNA molecule, wherein:
the RNA molecule encodes one or more reprogramming factor(s) selected from the group consisting of Oct4 protein, Sox2 protein, Klf4 protein, c-Myc protein, 1-Myc protein, Tert protein, Nanog protein, and Lin28 protein,
the transfecting results in the cell expressing the one or more reprogramming factor(s) which reprograms the cell to a less differentiated state, and
step (c) is performed without using irradiated human neonatal fibroblast feeder cells and occurs in the presence of a feeder cell conditioned medium.
2. The method of claim 1 , wherein the non-pluripotent cell is derived from a biopsy.
3. The method of claim 1 , wherein the non-pluripotent cell is harvested from a human subject.
4. The method of claim 2 , wherein the non-pluripotent cell is harvested from a dermal punch biopsy sample.
5. The method of claim 1 , wherein the non-pluripotent cell is a skin cell.
6. The method of claim 1 , further comprising contacting the cell with at least one member of the group: poly-L-lysine, poly-L-ornithine, RGD peptide, fibronectin, vitronectin, collagen, and laminin.
7. The method of claim 1 , wherein the synthetic RNA molecule contains at least one of a pseudouridine or a 5-methylcytidine residue.
8. The method of claim 1 , wherein the reprogramming medium is substantially free of immunosuppressants.