Three-Dimensional Earley Retinal Progenitor 3d Tissue Constructs Derived From Human Embryonic Stem Cell

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DEPARTMENT OF ANATOMY & NEUROBIOLOGY, REEVE-IRVINE RESEARCH CENTER, SUE AND BILL GROSS STEM CELL RESEARCH CENTER, UNIVERSITY OF CALIFORNIA AT IRVINE, IRVINE, CA, UNITED STATEST

Abstract

Purpose

To develop three-dimensional (3D) constructs of retinal pigment epithelium (RPE) and early retina progenitor cells from human embryonic stem cells (hESCs).

Methods

3D tissue constructs were developed by culturing hESC-derived neural retinal progenitors in a matrix on top of hESC-derived RPE cells in a cell culture insert. An osmolarity gradient maintained the nutrition of the 3D cell constructs. Cross-sections through hESC-derived tissue constructs were characterized by immunohistochemistry for various transcription factors and cell markers.

Results

hESC-derived tissue constructs expressed transcription factors characteristic of retinal development, such as pax6, Otx2, Chx10, retinal RAX; Brn3b (necessary for differentiation of retinal ganglion cells); and crx and nrl (role in photoreceptor development). Many cells expressed neuronal markers including nestin, beta-tubulin and microtubule-associated proteins.

Conclusions

This study shows for the first time that 3D early retinal progenitor tissue constructs can be derived from hESCs.

Keywords: Human embryonic stem cells; Retinal pigmented epithelium; Photoreceptor; Retinal progenitor differentiation; Three-dimensional culture

Abbreviations: hESC, human embryonic stem cells; RPE, retinal pigment epithelium; MAP, microtubule-associated protein; NF, neurofilament; Dkk, Dickkopf.

Gabriel Nistor, Magdalene J. Seiler, Fengrong Yana, David Fergusona and Hans S. Keirstead

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