Products for transfection and reprogramming

• US 9,127,248 B2
• Filed: 06/28/2013
• Issued: 09/08/2015
• Est. Priority Date: 12/05/2011
• Status: Active Grant

• Current Assignee: Factor Bioscience Inc
• Original Assignee: Factor Bioscience Inc
• Inventors: Angel, Matthew, Rohde, Christopher
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US13/931,251
• Publication Number:
  US 20130344602A1
• Time in Patent Office: 802 Days
• US Class Current: 1/1
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/33 other than alpha-amino carboxylic acids, e.g. beta-amino acids, taurine
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 Klf-4
• CPC Class Codes: C12N 2501/606 : c-Myc
• CPC Class Codes: C12N 2501/608 : Lin28
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/09 : from epidermal cells, from skin cells, from oral mucosa cells
• CPC Class Codes: C12N 2506/45 : from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0018 : Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0625 : Epidermal cells, skin cells; Cells of the oral mucosa
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0656 : Adult fibroblasts
• CPC Class Codes: C12N 5/0657 : Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0695 : Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12N 5/0696 : Artificially induced pluripotent stem cells, e.g. iPS

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.

Claims

1. A cell-culture medium comprising:

DMEM/F12, 10 μ

g/mL insulin, 5.5 μ

g/mL transferrin, 6.7 ng/mL sodium selenite, 20 ng/mL bFGF, and 5 mg/mL albumin, wherein the albumin is treated with an ion-exchange resin and/or charcoal.

2. The cell-culture medium of claim 1, further comprising at least one of:

15 mM HEPES, 2 mM L-alanyl-L-glutamine, 2 μ

g/mL ethanolamine, 4.5 μ

g/mL cholesterol, 25 μ

g/mL polyoxyethylenesorbitan monooleate, 2 μ

g/mL D-alpha-tocopherol acetate, and 1 μ

g/mL L-ascorbic acid 2-phosphate sequimagnesium salt hydrate.

3. The cell-culture medium of claim 1, wherein the albumin is treated with sodium octanoate.

4. The cell-culture medium of claim 1, wherein the albumin is brought to a temperature of at least 40°

C.

5. The cell-culture medium of claim 1, wherein the albumin is recombinant.

9. A kit comprising the cell-culture medium of claim 1.

10. The kit of claim 9, wherein the albumin is recombinant.

11. The kit of claim 9, further comprising one or more synthetic RNA molecules, wherein the one or more synthetic RNA molecules comprise at least one RNA molecule encoding one or more reprogramming factors.

12. The kit of claim 11, wherein the one or more synthetic RNA molecules comprise at least one RNA molecule encoding at least one of:

Oct4 protein, Sox2 protein, Klf4 protein, and c-Myc protein.

13. The kit of claim 11, wherein the one or more synthetic RNA molecules comprise at least one RNA molecule encoding Oct4 protein, at least one RNA molecule encoding Sox2 protein, at least one RNA molecule encoding Klf4 protein, and at least one molecule encoding c-Myc protein.

14. The kit of claim 11, wherein the one or more synthetic RNA molecules comprise at least one of:

a pseudouridine residue and a 5-methylcytidine residue.

6. A cell-culture medium comprising:

DMEM/F12, 10 μ

g/mL insulin, 5.5 μ

g/mL transferrin, 6.7 ng/mL sodium selenite, 20 ng/mL bFGF, and 5 mg/mL albumin, wherein less than 0.65% of the albumin'"'"'s dry weight comprises lipids and/or less than 0.35% of the albumin'"'"'s dry weight comprises free fatty acids.

7. The cell-culture medium of claim 6, further comprising at least one of:

15 mM HEPES, 2 mM L-alanyl-L-glutamine, 2 μ

g/mL ethanolamine, 4.5 μ

g/mL cholesterol, 25 μ

g/mL polyoxyethylenesorbitan monooleate, 2 μ

g/mL D-alpha-tocopherol acetate, and 1 μ

g/mL L-ascorbic acid 2-phosphate sequimagnesium salt hydrate.

8. The cell-culture medium of claim 6, wherein the albumin is recombinant.