Methods and products for transfection

Patent metadata

Field	Value
Publication number	US-9695401-B2
Application number	US-201615358818-A
Country	US
Kind code	B2
Filing date	Nov 22, 2016
Priority date	Dec 5, 2011
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Factor Bioscience Inc

Inventors

• Angel Matthew
• Rohde Christopher

Classifications

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P37/04

  Immunostimulants · CPC title

• A61P7/06

  Antianaemics · CPC title