Abstract
Purpose:
Despite intensive research on the potential use of retinal progenitor cells in the treatment of degenerative eye diseases, the definitive markers for these prognitors are still lacking. Our current study focused on CD117 (c-kit), a cell surface marker for haematopoietic stem cells and progenitor cells. Retinal progenitor cells were isolated from human fetus and the c-kit + and SSEA4- retinal cells were sorted out for further biological characterization
Methods:
hRPCS were isolated from human retina of 10 to 16 weeks gestational age (GA). Immunohistochemical staining was performed using antibody against c-kit + to determine the distribution of c-kit+/SSEA-4- cell in the eyes of fetus. The c-kit + / SSEA4- retinal progenitor cells sorted out by flow cytometry were subjected to cell-mediated immunity fluorescent and cell cycle analysis for their capablity to proliferation and differentiation.
Results:
A few c-kit + / SSEA4- cells were detected in the inner part of fetal retina. Sorted c-kit + / SSEA4- cells expressed some retinal stem cell markers including Pax6, Sox2 and Nestin. Over 80% of the cells expressed ki67, and cell cycle analysis demonstrated that more than 40% of the cells were in their proliferation phase. These results clearly demonstrate proliferation property of these cells. When cultured in differentiation medium, these cells expressed markers found in photoreceptor cells and glial cells such as CRX, recoverin and GFAP.
Conclusions:
c-kit can be used as a surface marker for retinal progenitor cells. The c-kit + / SSEA4- retinal progenitor cell isolated fetal eyes exhibit the ability to self-renew and differentiate into retinal cells.
Keywords: 721 stem cells •
497 development •
688 retina