Abstract
Purpose :
Ocular neural crest cells give rise to multiple tissue types in the anterior segment of the eye during embryogenesis. Nevertheless, it is challenging to isolate and propagate human ocular neural crest stem cells (oNCSCs) in culture. We aim to characterize iPSC-derived oNCSCs and their potential as a renewable source of corneal endothelial cells as well as other type of cells of the anterior segment of the eye.
Methods :
The induction of oNCSC from human pluripotent stem cell (hPSC) was driven by small molecules in a stepwise fashion. Under differentiation conditions, neural crest cells were directed towards differentiating into anterior segment cells such as corneal endothelial-like cells. Cell morphology, differentiation and the expression of lineage specific markers were analyzed.
Results :
Ocular neural crest cells were derived from human iPSCs via eye field lineage specification. Initially, hPSCs were converted into eye field stem cells (EFSCs), which subsequently were induced to become oNCSCs. Their identity was confirmed by the expression of typical eye field transcription factors such as PAX6 and LHX2. The induced oNCSCs expressed gene markers such as p75NTR, HNK-1, NESTIN, DCX and NCAM., and were able to differentiate into corneal endothelial-like cells and other types of anterior segment cells in culture. They also spontaneously formed distinguishable subtypes such as stem cell-like clusters and migratory cells.
Conclusions :
This study provides a useful system for studying ocular neural crest cell biology and anterior segment cell fate specification in vitro. This study also offers a potentially renewable source of anterior segment cells that could have valuable clinical applications.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.