Modalities for the treatment of degenerative diseases of the retina

Patent metadata

Field	Value
Publication number	US-9730962-B2
Application number	US-201514692191-A
Country	US
Kind code	B2
Filing date	Apr 21, 2015
Priority date	Jan 23, 2004
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

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.

First claim

We claim:

1. A method of generating a retinal pigment epithelium (RPE) cell preparation suitable for transplantation therapy comprising:
(a) obtaining RPE cells that have a cobblestone polygonal epithelial-like cellular morphology and pigmentation, wherein the RPE cells include cells that express RPE65 and bestrophin;
(b) dispersing the RPE cells and culturing the dispersed RPE cells under adherent conditions in the presence of basic FGF, wherein the cultured RPE cells dedifferentiate and proliferate, losing pigmentation and epithelial-like morphology;
(c) culturing the dedifferentiated cells in the absence of basic FGF, wherein the cells form an RPE cell monolayer, become quiescent, and regain pigmentation and epithelial-like morphology, and wherein the RPE cell monolayer includes cells that express RPE65 and bestrophin; and
(d) expanding the RPE cells by repeating steps (b) and (c) through multiple passages.

2. The method of claim 1 , wherein steps (b) and (c) are repeated to provide for three passages.

3. The method of claim 1 , wherein the RPE cell preparation is a human RPE cell preparation.

4. The method of claim 1 , wherein the dedifferentiated cells that have lost pigmentation and epithelial morphology are positive for mitf, Pax6, tubulin beta III and nestin.

5. The method of claim 1 , wherein the RPE cell monolayer includes cells that are Pax6−, bestrophin+, CRALBP+, PEDF+, and express RPE65.

6. The method of claim 1 , wherein the RPE cell monolayer is characterized by the absence of any cells expressing at least one ES cell marker selected from the group consisting of Oct4, Sox2, and TDGF-1.

7. The method of claim 1 , further comprising
(e) isolating a pure preparation of human RPE cells suitable for transplantation therapy after the multiple passages of (d).

8. The method of claim 7 , wherein the preparation of RPE cells suitable for transplantation therapy is provided with a matrix or substrate.

9. The method of claim 7 , wherein the preparation of RPE cells suitable for transplantation therapy is provided in a suspension.

Assignees

• Ocata Therapeutics Inc
• Astellas Inst For Regenerative Medicine

Inventors

• Klimanskaya Irina V
• Lanza Robert P

Classifications

• 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

• A61P9/14

  Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P27/02

  Ophthalmic agents · CPC title

• A61P27/00

  Drugs for disorders of the senses · CPC title