Journal
ACTA OPHTHALMOLOGICA
Volume 89, Issue 8, Pages e635-e644Publisher
WILEY
DOI: 10.1111/j.1755-3768.2011.02198.x
Keywords
adult human; ciliary body; iris pigment epithelium; peripheral iridectomy; progenitor cell; retinal stem cell
Categories
Funding
- The Medical Technical Department, OUH
- Center for Eye Research, OUH
- Research Council of Norway
- Norwegian Association of the Blind and Partially Sighted, Blindemisjonen IL
- Faculty of Medicine University of Oslo
- Oslo University Hospital
- South-Eastern Norway Regional Health Authority
- Hungarian Scientific Research Fund [OTKA NI 67877, K 61868]
- Hungarian Ministry of Health
- EU [MRTNCT-2006-036032, MRTN-CT 2006-035624, LSHB-CT-2007-037730, TAMOP 4.2.1./B-09/1/KONV-2010-0007]
- New Hungary Development Plan
- European Social Fund
- Pfizer Research Partnership Agreement (Hungary)
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Purpose: The identification of cells with properties of retinal progenitor cells (RPCs) in the adult human ciliary margin (CM) prompted a number of studies of their proliferative and differentiation potential. One of the remaining challenges is to find a feasible method of isolating RPCs from the patients eye. In the human CM, only the iris pigment epithelium (IPE) is easily obtained by a minimally invasive procedure. In the light of recent studies questioning the existence of RPCs in the adult mammalian CM, we wanted to assess the potential of the adult human IPE as source of RPCs. Methods: The IPE were isolated from peripheral iridectomies during glaucoma surgery, and IPE and ciliary body (CB) epithelium were also isolated from post-mortem tissue. Cells were cultivated in sphere-promoting conditions or as monolayers. Whole-tissue samples, undifferentiated and differentiated cells were studied by immunocytochemistry, RT-PCR and transmission electron microscopy. Results: The adult human IPE, like the CB, expressed markers of RPCs such as Pax6, Sox2 and Nestin in vivo. Both sphere-promoting and monolayer cultures preserved this phenotype. However, both IPE/CB cultures expressed markers of differentiated epithelial cells such as Claudin, microphtalmia-associated transcription factor (MITF) and Cytokeratin-19. Ultrastructurally, IPE spheres displayed epithelial-like junctions and contained mature melanosomes. After induced differentiation, IPE-derived cells showed only partial neuronal differentiation expressing beta-III-tubulin, Map-2 and Rhodopsin, whereas no mature glial markers were found. Conclusion: Proliferative cells with some properties of RPCs can be isolated from the adult human IPE by peripheral iridectomies. Yet, many cells retain properties of differentiated epithelial cells and lack central properties of somatic stem cells.
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