Methods of Producing RPE Cells and Compositions of RPE Cells

Abstract

The present invention provides improved methods for producing RPE cells from human embryonic stem cells or from other human pluripotent stem cells. The invention also relates to human retinal pigmented epithelial cells derived from human embryonic stem cells or other human multipotent or pluripotent stem cells. hRPE cells derived from embryonic stem cells are molecularly distinct from adult and fetal-derived RPE cells, and are also distinct from embryonic stem cells. The hRPE cells described herein are useful for treating retinal degenerative diseases.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of priority to U.S. provisional application Nos. 60 / 998,766, filed Oct. 12, 2007, 60 / 998,668, filed Oct. 12, 2007, 61 / 009,908, filed Jan. 2, 2008, and 61 / 009,911, filed Jan. 2, 2008. The disclosures of each of the foregoing applications are hereby incorporated by reference in their entirety.BACKGROUND OF THE INVENTION[0002]The retinal pigment epithelium (RPE) is the pigmented cell layer just outside the neurosensory retina. This layer of cells nourishes retinal visual cells, and is attached to the underlying choroid (the layer of blood vessels behind the retina) and overlying retinal visual cells. The RPE acts as a filter to determine what nutrients reach the retina from the choroid. Additionally, the RPE provides insulation between the retina and the choroid. Breakdown of the RPE interferes with the metabolism of the retina, causing thinning of the retina. Thinning of the retina can have serious consequences. For example...

AI Technical Summary

Benefits of technology

[0066]In certain embodiments of any of the foregoing, said substantially purified culture of RPE cells (with or without mature RPE cells) are frozen for storage. The cells may be stored by any appropriate method known in the art, e.g., cryogenically frozen and may be frozen at any temperature appropriate for storage of the cells. For example, the cells may be frozen at approximately −20° C., −80° C., −120° C., or at any other temperature appropriate for storage of cells. Cryogenically frozen cells are stored in appropriate containers and prepared for storage to reduce risk of cell damage and maximize the likelihood that the cells will survive thawing. In other embodiments, RPE cells can be maintained at room temperature, or refrigerated at, for example, approximately 4° C.

[0067]In certain embodiments of any of the foregoing, human RPE cells are produced in accordance with Good Manufacturing Practices (GMP). In certain embodiments of any of the foregoing, the human embryonic stem cells from which the RPE cells are differentiated were derived in accordance with Good Manufacturing Practices (GMP). In certain embodiments of any of the foregoing, the human embryonic stem cells from which the RPE cells are differentiated were derived from one or more blastomeres removed from an early stage embryo without destroying the remaining embryo. As such, the invention provides GMP compliant preparations of RPE cells, including substantially purified preparations of RPE cells. Such preparations are substantially free of viral, bacterial, and / or fungal contamination or infection.

Problems solved by technology

Breakdown of the RPE interferes with the metabolism of the retina, causing thinning of the retina.

Thinning of the retina can have serious consequences.

For example, thinning of the retina may cause “dry” macular degeneration and may also lead to the inappropriate blood vessel formation that can cause “wet” macular degeneration).

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