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R. S. Kolberg, K. Haug, E. Gulliksen, C. Sandberg, I. A. Langmoen, B. Nicolaissen, M. C. Moe; Comparison of Progenitor Cells Derived From the Adult Human Iris and Ciliary Body. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5151.
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We have recently shown that spheres derived from the ciliary body (CB) contain progenitor cells with a mixed epithelial and neural phenotype compared to CNS-neurospheres. Next we wanted to study the potential for neural differentiation of progenitors isolated from both the adult human CB and the iris pigmented epithelium (IPE), with the purpose of determining their future potential in cell-based therapy of retinal disease.
The CB and IPE were dissected from donor eyes, and IPE was also isolated from peripheral iridectomies. The tissue was dissociated into single cells and grown as spheres for 2-5 passages. To differentiate the cells, spheres were plated on an adhesive substrate and cultured for 3 weeks. Fixed cells were studied with immunocytochemistry, electron microscopy and molecular biology.
After 1 week, pigmented spheres of at least 100µm were found in all cultures from post mortem IPE and CB (n=18), but only in 2 out of 12 cultures from peripheral iridectomies. IPE-spheres were on average more heavily pigmented than CB-spheres, but gradual loss of pigment was observed on repeated passaging. At P2, 38% of cells in CB spheres were positive for nestin, 41% for Ki67, and 42% for claudin-1. In the IPE-spheres 42% of cells were positive for nestin, 23% for Ki-67, and 38% for claudin-1. GFAP-staining was not detected. Upon induced differentiation, cells from both CB and IPE spheres (P2-4) acquired a mature neuronal morphology and strongly expressed beta-III-tubulin and Map-2. A subpopulation of the beta-III-tubulin-positive cells with round cell soma and short extensions also showed robust expression of rhodopsin.
The human iris harbours sphere-forming cells with similar properties to those of the CB, including the ability to proliferate in vitro for several passages and differentiate into cells positive for markers of mature neurons and photoreceptors.
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