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THREE-DIMENSIONAL
EARLEY RETINAL PROGENITOR 3D TISSUE CONSTRUCTS DERIVED
FROM HUMAN EMBRYONIC STEM CELL
Gabriel
Nistor, Magdalene J. Seiler, Fengrong Yana, David Fergusona
and Hans S. Keirstead |
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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 States
Received 18 May 2009;
revised 22 April 2010;
accepted 26 April 2010.
Available online 4 May 2010. |
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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. |
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