Three-dimensional early retinal progenitor 3D tissue constructs derived from human embryonic stem cells.
OBJECTIVE
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.
Nistor
Gabriel
G
Department of Anatomy & Neurobiology, Reeve-Irvine Research Center, Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA, USA.
Seiler
Magdalene J
MJ
Yan
Fengrong
F
Ferguson
David
D
Keirstead
Hans S
HS
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000502:physiology*; D005109:Extracellular Matrix; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000502:physiology; D012160:Retina / Q000502:physiology*; D055213:Retinal Pigment Epithelium / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014157:Transcription Factors / Q000378:metabolism
D014157:Transcription Factors