# US 9,040,039 B2 · Modalities for the treatment of degenerative diseases of the retina

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- Number: US 9,040,039 B2
- Document ID: US9040039B2
- Status: Granted
- Kind: B2
- Application number: 13/865,751
- Filed: 2013-04-18
- Published: 2013-11-14
- Granted: 2015-05-26
- Earliest claimed: 2004-01-23
- Assignees: Ocata Therapeutics, Inc.
- Applicants: Ocata Therapeutics, Inc.
- Inventors: Irina V. Klimanskaya; Robert P. Lanza
- Web page: https://patentgrep.com/patent/US9040039B2

## CPC

- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- 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)
- A61K 35/30: Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
- A61K 35/36: Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells (islets of Langerhans A61K35/39)
- A61K 35/44: Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
- A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
- A61P 25/16: Anti-Parkinson drugs
- A61P 27/02: Ophthalmic agents
- A61P 27/06: Antiglaucoma agents or miotics
- C12N 5/062: {Sensory transducers, e.g. photoreceptors; Sensory neurons, e.g. for hearing, taste, smell, pH, touch, temperature, pain}
- C12N 2500/32: Amino acids
- C12N 2500/44: Thiols, e.g. mercaptoethanol
- C12N 2501/01: Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/15: Transforming growth factor beta (TGF-β)
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2501/235: Leukemia inhibitory factor [LIF]
- C12N 2501/33: Insulin
- C12N 2501/60: Transcription factors
- C12N 2506/02: from embryonic cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

This invention relates to methods for improved cell-based therapies for retinal degeneration and for differentiating human embryonic stem cells and human embryo-derived into retinal pigment epithelium (RPE) cells and other retinal progenitor cells.

## Claims

### Claim 1

A method for the differentiation of human embryonic stem (hES) cells, said method comprising:
a) culturing a multilayer of cells, produced from hES cells, for a sufficient time for the appearance of pigmented cells comprising brown pigment dispersed in their cytoplasm, under conditions that do not maintain stem cells in an undifferentiated state, wherein the hES cells are stem cells that express Oct-4, alkaline phosphatase, SSEA-3, SSEA-4, TRA-I-60 and TRA-I-81; and
b) isolating and culturing the pigmented cells under adherent conditions to form a population of cells that expresses bestrophin, CRALBP, PEDF1 and RPE65.

### Claim 2 (depends on 1)

The method of claim 1, wherein the population of cells in (b) comprises cells having a cobblestone, polygonal appearance.

### Claim 3 (depends on 2)

The method of claim 2, wherein the population of cells in (b) comprises cells that are tightly packed.

### Claim 4 (depends on 1)

The method of claim 1, further comprising isolating the population of cells in (b) as a suspension.

### Claim 5 (depends on 1)

The method of claim 1, further comprising isolating the population of cells in (b) as a monolayer.

### Claim 6 (depends on 1)

The method of claim 1, further comprising isolating and placing the population of cells in (b) on a matrix or a substrate.

### Claim 7 (depends on 1)

The method of claim 1, wherein the multilayer of cells is produced by allowing hES cells to overgrow and form the multilayer of cells.

### Claim 8 (depends on 1)

The method of claim 1, wherein isolating the pigmented cells comprises contacting the pigmented cells with one or more enzymes selected from the group consisting of trypsin, collagenase and dispase.

### Claim 9 (depends on 1)

The method of claim 1, further comprising isolating and transplanting the population of cells into an eye of a patient to treat a retinal disease or condition.

### Claim 10 (depends on 9)

The method of claim 9, wherein the population of cells is transplanted by vitrectomy surgery into the subretinal space of the eye of the patient.

### Claim 11 (depends on 9)

The method of claim 9, wherein the patient has a retinal disease or condition selected from the group consisting of retinitis pigmentosa, retinal detachment, retinal dysplasia, retinal atrophy, retinopathy, macular dystrophy, cone dystrophy, cone-rod dystrophy, Malattia Leventinese, Doyne honeycomb dystrophy, Sorsby's dystrophy, pattern/butterfly dystrophies, Best vitelliform dystrophy, North Carolina dystrophy, central areolar choroidal dystrophy, angioid streaks, or a toxic maculopathy.

### Claim 12 (depends on 9)

The method of claim 9, wherein the patient has Stargardt disease, pathologic myopia, retinitis pigmentosa, or macular degeneration.

### Claim 13 (depends on 9)

The method of claim 9, wherein the patient has age-related macular degeneration.

### Claim 14 (depends on 9)

The method of claim 9, wherein the population of cells is cryopreserved and then thawed prior to being transplanted into the patient.

### Claim 15 (depends on 9)

The method of claim 9, wherein the population of cells is in a suspension.

### Claim 16 (depends on 9)

The method of claim 9, wherein the population of cells is a monolayer.

### Claim 17 (depends on 9)

The method of claim 9, wherein the population of cells is on a matrix or a substrate.