23 Jul Inducing method for directionally differentiating human embryonic stem cells to corneal endothelial cells
Abstract
The invention discloses an inducing method for directionally differentiating human embryonic stem cells to corneal endothelial cells. The method comprises the steps of: cultivating the human embryonic stem cells on a mouse embryonic fibroblast feed layer; sorting human embryonic stem cell clone groups in good state; grafting the groups on a human corneal stromal fibroblast layer processed by mitomycin C and cultivating for 7 days, wherein the human embryonic stem cells are differentiated to rosettes; separating and transferring the rosettes from the human corneal stromal fibroblast layer to a culture bottle; cultivating continuously for 7 days by using a neural crest stem cell culture medium; sorting the neural crest stem cells by a flow cytometry; adding the neural crest stem cells into the culture bottle; placing in a 5% CO2 incubator for incubating and cultivating at 37 DEG C by using a human corneal endothelial cell culture medium; changing the liquid every other day; and cultivating for about 10 days to obtain the corneal endothelial cells. The multiplication capacity of the corneal endothelial cells are similar to that of human corneal endothelial cells and the corneal endothelial cells can be transferred to 1-2 generations in vitro maximally. The corneal endothelial cells can be used as seed cells for cornea construction and transplant in tissue engineering.
Year: 2013
Country: CN
Doc No: 102952777