Abstract
Abstract: :
Purpose:To examine if and where stem cells are present in the normal human eye with immunohistochemistry Methods: Human cadaver eyes (N=12) were collected from eye bank. Eyes were fixed in 1% formaldehyde. 6µ paraffin sections were cut on a cryostat from embedded samples.The sections were deparaffinized with a series of washes. Slides were then exposed to primary antibodies of Nestin, Notch, and Chx–10. After overnight incubation at 4oC, slides were incubated with conjugated secondary antibodies. DAB chromagen was added to visualize the peroxidase–conjugated second antibody. Cell nuclei were stained with DAPI during the last rinse. Slides were mounted, and examined using a Zeiss Axioscop 2. Results: Positive cells were most prominent in the nonpigmented ciliary epithelium, iris epithelium and some areas of retinal pigment epithelium. Only cells that colocalized all three antibodies immunohistochemically were considered positive. Some amount of autofluorescence was noted inside the vasculature with red blood cells. Conclusions: Stem cells were predominantly present within the iris epithelium and non–pigmented ciliary epithelium of normal human eyes analogous to ciliary ridge of other species
Keywords: immunohistochemistry • detection • retinal development