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Yoshinori Nakai, Morio Ueno, Hiroshi Tanaka, Makoto Fukuta, Makoto Ikeya, Junya Toguchida, Shigeru Kinoshita; Induction of corneal endothelial cells from human pluripotent stem cells through derived neural crest like cells.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):526.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal endothelium, corneal keratocytes and trabecular meshwork cells, which construct anterior segment of the eye, are developed from neural crest cells. We inducted corneal endothelium like cells used neural crest cells derived from human pluripotent cells, embryonic stem (ES) cells and induced pluripotent stem (iPS) cells. This study involves possibility of producing cell sources for regenerative medicine with high efficiency.
ES/iPS cells are isolated from the feeder cells and they are cultured on Matrigel in serum-free medium. The proportion of CD271(p75NTR) high positive cells are sorted by using flow cytometory, as neural crest cells. Then they cultured with corneal endothelium conditioned medium for one week. We analyzed them by use of immunostaining and quantitative polymerase chain reaction (qPCR).
Neural crest cells derived from human ES/iPS cells proliferate in the conditioned medium and they changed their morphology epithelial like cells, immunostaining testing showed Zo-1 positive cells. The induced cells express gene markers for the most abundant genes of corneal endothelium, Na/K ATPase alpah1 with PCR.
This study provides a highly practical system for inducing corneal endothelium from human ES/iPS cells through derived neural crest cells.
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