Compound screening using cardiomyocytes

• US 7,851,167 B2
• Filed: 09/22/2008
• Issued: 12/14/2010
• Est. Priority Date: 07/12/2001
• Status: Expired due to Fees

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Geron Corporation
• Inventors: Xu, Chunhui
• Primary Examiner(s): Ton; Thaian N
• Application Number:
  US12/234,916
• Publication Number:
  US 20090017465A1
• Time in Patent Office: 813 Days
• Field of Search: 435/7.21, 435/325, 435/363, 435/366, 435/377
• US Class Current: 435/7.21
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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 2500/30 Organic components metal chelators C12N2500/10; calcitonin C12N2500/14; transferrin C12N2500/24
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2500/62 DMSO
• CPC Class Codes: C12N 2501/06 Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells

Abstract

This invention provides populations human cells of the cardiomyocyte lineage. The cells are obtained by causing cultures of pluripotent stem cells to differentiate in vitro, and then harvesting cells with certain phenotypic features. Differentiated cells bear cell surface and morphologic markers characteristic of cardiomyocytes, and a proportion of them undergo spontaneous periodic contraction. Highly enriched populations of cardiomyocytes and their replicating precursors can be obtained, suitable for use in a variety of applications, such as drug screening and therapy for cardiac disease.

Claims

1. A method of screening a compound for an effect on a cardiomyocyte in cell culture, comprising:

a) contacting the cell culture with the compound; and

b) observing a change in a cardiomyocyte in the cell culture compared to a cardiomyocyte in an untreated cell culture and

c) determining a difference between the treated and untreated cardiomyocyte in the respective cell cultures,

wherein the cell culture comprises 5% or more cardiomyocytes that are at least 99.6% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

2. The method of claim 1, wherein the compound is a drug.

3. The method of claim 2, wherein the drug is calcium channel blocker.

4. The method of claim 2, wherein the drug is a β

-adrenoreceptor agonist.

5. The method of claim 2, wherein the drug is an α

-adrenoreceptor agonist.

6. The method of claim 1, wherein the compound is a peptide.

7. The method of claim 1, wherein the compound is an oligonucleotide.

8. The method of claim 1, wherein the compound is a cytotoxin.

9. The method of claim 1, wherein the cell culture comprises 5% or more cardiomyocytes that are 99.7% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

10. The method of claim 1, wherein the cell culture comprises 5% or more cardiomyocytes that are 99.8% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.38.

11. The method of claim 1, wherein the cell culture comprises 5% or more cardiomyocytes that are 99.9% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.38.

12. The method of claim 1, wherein the cell culture comprises 5% or more cardiomyocytes that have the same genome as an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

13. A method of screening a compound for an effect on a cell in cell culture, comprising:

a) contacting the cell culture with a compound; and

b) observing a change in a myosin heavy chain expressing cell in the cell culture compared to a myosin heavy chain expressing cell in an untreated cell culture and

c) determining a difference between the treated and untreated myosin heavy chain expressing cell in the respective cell cultures,

wherein the cell culture comprises 5% or more cells that express myosin heavy chain and are at least 99.6% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

14. The method of claim 13 wherein the compound is a drug.

15. The method of claim 14, wherein the drug is calcium channel blocker.

16. The method of claim 14, wherein the drug is a β

-adrenoreceptor agonist.

17. The method of claim 14, wherein the drug is an α

-adrenoreceptor agonist.

18. The method of claim 13, wherein the compound is a peptide.

19. The method of claim 13, wherein the compound is an oligonucleotide.

20. The method of claim 13, wherein the compound is a cytotoxin.

21. The method of claim 13, wherein the cell culture comprises 5% or more cardiomyocytes that are 99.7% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

22. The method of claim 13 wherein the cell culture comprises 5% or more cardiomyocytes that are 99.8% identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

23. The method of claim 13, wherein the cell culture comprises 5% or more cardiomyocytes that are 99.9% genetically identical to an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.

24. The method of claim 13, wherein the cell culture comprises 5% or more cardiomyocytes that have the same genome as an established line of non-malignant cells which express SSEA3, SSEA4, Tra-1-60 and Tra-1-81.