In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells

Fernando H Lojudice,Francesco Innocenti,Maurício Maia,Rodrigo A Brant Fernandes,Mari C Sogayar,Priscila Cristovam,Carlos E Franciozi,Rubens Belfort

doi:10.1186/s40942-019-0194-7

Abstract

BackgroundThe World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most macular degeneration, i.e., age-related macular degeneration (AMD) develops in the elderly; however, certain hereditary diseases, such as the Stargardt disease, also affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). In wet AMD, numerous drugs are available to successful treat the disease; however, no proven therapy currently is available to treat dry AMD or Stargardt. Since its discovery, human embryonic stem cells (hESCs) have been considered a valuable therapeutic tool. Some evidence has shown that transplantation of RPEs differentiated from hESCs cells can result in recovery of both RPE and photoreceptors and prevent visual loss.MethodsThe human embryonic WA-09 stem cell lineage was cultured under current Good Manufacturing Practices (cGMP) conditions using serum-free media and supplements. The colonies were isolated manually and allowed to spontaneously differentiate into RPE cells.ResultsThis simple and effective protocol required minimal manipulation and yielded more than 10e8 RPE cells by the end of the differentiation and enrichment processes, with cells exhibiting a cobblestone morphology and displaying cellular markers and a gene expression profile typical of mature RPE cells. Moreover, the differentiated cells displayed phagocytic activity and only a small percentage of the total cells remained positive for the Octamer-binding transcriptions factor 4 (OCT-4) pluripotency cell marker.ConclusionsThese results showed that functional RPE cells can be produced efficiently and suggested the possibility of scaling-up to aim at therapeutic protocols for retinal diseases associated with RPE degeneration.

Highlights

The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020
Age-related macular degeneration (AMD) is a leading cause of irreversible blindness worldwide [1, 2] that has been estimated to affect more than 8 million people in the United States alone
The advanced forms of age-related macular degeneration (AMD) are neovascular and non-neovascular atrophic AMD, both of which are associated with visual acuity loss [5]

Summary

Introduction

The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. I.e., age-related macular degeneration (AMD) develops in the elderly; certain hereditary diseases, such as the Stargardt disease, affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). Age-related macular degeneration (AMD) is a leading cause of irreversible blindness worldwide [1, 2] that has been estimated to affect more than 8 million people in the United States alone. Outer retinal degenerative diseases, such as AMD, lead to progressive, irreversible loss of the central visual acuity, with the retinal pigmented epithelium (RPE) the focus of the disease pathophysiology. The advanced forms of AMD are neovascular (wet AMD) and non-neovascular atrophic AMD (dry AMD), both of which are associated with visual acuity loss [5].

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