IP Library Granted Patent US 9,423,394
Granted Patent B2
US 9,423,394 · App. 14/106,137 · Granted Aug 23, 2016

Retinal pigment epithelial primary cell culture system producing subcellular deposits

Inventors: Christine A. Curcio (Birmingham, AL); Clyde Guidry (Birmingham, AL)
G01N33/5058C12N5/0621G01N33/5035
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Quick Facts
Patent No.
US 9,423,394
App. No.
14/106,137
Granted
Aug 23, 2016
Kind
B2
Abstract

The present invention provides a retinal pigment epithelial (RPE) primary cell culture system on a material surface seeded at a high density that produces a layer of subcellular deposits, wherein the RPE primary cells are harvested from non-fetal tissue. The present invention additionally provides methods of making and using the PRE primary cell culture system.

Claims (34)

1. A retinal pigment epithelial (RPE) primary cell culture system, comprising:

RPE primary cells seeded at a high density on a material surface and cultured without subculturing, wherein the RPE primary cells are harvested from non-fetal tissue; and

a confluent layer of lipid-rich subcellular deposits between the RPE primary cells and the material surface.

2. The RPE primary cell culture system of claim 1 , wherein the RPE cells of the cell culture system are contact inhibited and phenotype arrested.

3. The RPE primary cell culture system of claim 1 , wherein the cells in the RPE primary cell culture system are in a confluent monolayer on the material surface.

4. The RPE primary cell culture system of claim 1 , wherein the confluent layer of lipid-rich subcellular deposits comprises drusen and/or basal linear deposits.

5. The RPE primary cell culture system of claim 1 , wherein the non-fetal tissue is from a mammal, a bird, a reptile, an amphibian, and/or a fish.

6. The RPE primary cell culture system of claim 5 , wherein the mammal is a human, a non-human primate or a pig.

7. A method of identifying a test agent that modulates the amount of lipid-rich subcellular deposits produced in an RPE primary cell culture system, comprising:

contacting the RPE primary cell culture system of claim 1 with a test agent; and

determining the amount of lipid-rich subcellular deposits produced by the RPE primary cell culture system contacted with said test agent as compared to a control RPE primary cell culture system not contacted with said test agent, thereby identifying a test agent that modulates the amount of lipid-rich subcellular deposits produced by the RPE primary cell culture system.

8. The method of claim 7 , wherein the test agent modulates the amount of lipid-rich subcellular deposits by reducing the amount of, preventing the accumulation of, or preventing further accumulation of the lipid-rich subcellular deposits produced by the RPE primary cell culture system contacted with the test agent as compared to a control RPE primary cell culture system not contacted with the test agent.

9. The method of claim 8 , wherein the lipid-rich subcellular deposits comprise drusen and/or basal linear deposits (BlinD).

10. A method for identifying an agent for treating macular degeneration, comprising:

contacting the RPE primary cell culture system of claim 1 with an agent; and

determining the amount of lipid-rich subcellular deposits produced by the RPE primary cell culture system contacted with said agent as compared to a control RPE primary cell culture system not contacted with said agent,

wherein an agent that reduces the amount of, prevents the accumulation of, or prevents further accumulation of lipid-rich subcellular deposits produced by said RPE primary cell culture system contacted with said agent as compared to a control RPE primary cell culture system not contacted with said agent is identified as an agent for treating macular degeneration.

11. A method for identifying an agent for preventing macular degeneration, comprising:

contacting the RPE primary cell culture system of claim 1 with an agent; and

determining the amount of lipid-rich subcellular deposits produced by the RPE primary cell culture system contacted with said agent as compared to a control RPE primary cell culture system not contacted with said agent,

wherein an agent that reduces the amount of, prevents the accumulation of, or prevents further accumulation of lipid-rich subcellular deposits produced by said RPE primary cell culture system contacted with said agent as compared to a control RPE primary cell culture system not contacted with said agent is identified as an agent for preventing macular degeneration.

12. A method of producing a retinal pigment epithelial (RPE) primary cell culture system that produces a confluent layer of lipid-rich subcellular deposits, comprising:

(a) seeding RPE primary cells on a material surface at a high density, wherein said RPE primary cells are harvested from non-fetal tissue; and

(b) culturing the RPE primary cells seeded on said material surface without subculturing, thereby producing a RPE primary cell culture system that produces a confluent layer of lipid-rich subcellular deposits between the RPE primary cells and the material surface.

13. The method of claim 12 , wherein the RPE cells are seeded at a density of 40,000 cells to about 200,000 cells per square centimeter of material surface.

14. The method of claim 12 , wherein the confluent layer of lipid-rich subcellular deposits comprises drusen and/or basal linear deposits (BlinD).

15. The method of claim 12 , wherein the RPE primary cells are subjected to high density centrifugation prior to seeding on the material surface.

16. The method of claim 12 , wherein the RPE primary cells are from the eye of an organism excised from said organism 0 to 60 min post-mortem.

17. The method of claim 12 , wherein the cells in the RPE primary cell culture system are in a confluent monolayer on the material surface.

18. The method of claim 12 , wherein the non-fetal tissue is from an organism that is a mammal, a bird, a reptile, an amphibian, and/or a fish.

19. The method of claim 18 , wherein the mammal is a human, a non-human primate or a pig.

20. A method of producing a confluent layer of lipid-rich subcellular deposits from a retinal pigment epithelial (RPE) primary cell culture system, comprising:

(a) seeding RPE primary cells at a high density on a material support, wherein said RPE primary cells are harvested from non-fetal tissue; and

(b) culturing the RPE primary cells seeded on said material support without subculturing, thereby producing a confluent layer of lipid-rich subcellular deposits between the RPE primary cells and the material support.

Assignments (5)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 29, 2016
From: THERAOPTIX, INC.
To: CURCIO, CHRISTINE A; ROIZMAN, KEITH
Reel/Frame 037849/0888 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2014
From: UAB RESEARCH FOUNDATION
To: GUIDRY, CLYDE, DR.; CURCIO, CHRISTINE A., DR.
Reel/Frame 034159/0151 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2014
From: CURCIO, CHRISTINE A., DR.; GUIDRY, CLYDE, DR.
To: THERAOPTIX, INC.
Reel/Frame 034159/0548 →
CONFIRMATORY LICENSE Recorded Oct 1, 2014
From: UNIVERSITY OF ALABAMA AT BIRMINGHAM
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 033870/0749 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 26, 2014
From: CURCIO, CHRISTINE A.; GUIDRY, CLYDE
To: UAB RESEARCH FOUNDATION
Reel/Frame 032300/0607 →
Continuity (1)
Related Publication 20150168378A1 · Jun 18, 2015