Abstract
Purpose:
There is an increasing interest in generating retinal pigment epithelial (RPE) cells from stem cells for treatment of macular degeneration. Human sweat glands are a rich source of nestin-positive stem cells. In this study, the possibility of inducing RPE-specific markers in human sweat gland-derived stem cells (SGSCs) by xenogeneic co-culture with porcine RPE cells was investigated.
Methods:
SGSCs were isolated out of adult human scalp skin, purified and seeded on laminin-coated cover slips. Then they were co-cultured with porcine RPE cells seeded on laminin-coated transwell inserts or mono-cultured without RPE cells for 5 days. Afterwards they were washed, fixed, stained (Bestrophin, MITF, PMEL, MERTK, CRALBP and RPE65) and analyzed by fluorescence microscopy.
Results:
SGSCs expressed Bestrophin and MITF on laminin-coated cover slips even when mono-cultured without RPE cells whereas PMEL, MERTK, CRALBP and RPE65 were only detectable after co-culture with RPE cells.
Conclusions:
Adult human sweat gland-derived stem cells can be directed into expressing RPE-specific proteins even by xenogeneic co-culture with porcine RPE cells. The presented system is an effective tool to predict the behavior of SGSCs after transplantation into the subretinal environment by mimicking the situation in vivo.
Keywords: 412 age-related macular degeneration •
741 transplantation •
721 stem cells