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M. C. Moe, R. C. Frøen, E. Vik-Mo, C. Sandberg, H. Olstorn, I. A. Langmoen, B. Nicolaissen, University of Oslo, Norway; A Comparison of the in vitro Niche of Retinal and Brain Progenitor Cells From the Adult Human CNS. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3554.
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Retinal stem/progenitor cells present in the ciliary body (CB) of the adult human eye can be isolated and propagated as spheres in vitro and differentiated into retinal cell types. As these progenitor cells are isolated from a non-neural epithelium of neuroepithelial origin, it is vital to compare the CB spheres with other CNS neurospheres to characterize their neural and non-neural properties. We have for the first time compared CB spheres isolated from the adult human eye with neurospheres derived from the adult human subventricular zone (SVZ), which is considered the largest and best characterized stem cell niche in the CNS.
The pigmented CB was dissected from donor eyes (n=8). Biopsies from the ventricular wall were harvested during neurosurgery due to medical intractable epilepsy (n=7). Tissue from CB and SVZ were dissociated into single cells and cultivated in a defined sphere-promoting medium. Fixed spheres were studied by immunocytochemistry, quantitative RT-PCR and scanning/transmission electronmicroscopy.
The CB spheres showed increased growth compared to SVZ spheres. Genes involved in the proliferation rate of progenitor cells such as EGF/TGFβ receptors were upregulated in CB spheres compared to SVZ spheres. Markers of immature neural progenitor cells (nestin, sox-2, GFAP) were expressed in both CB - and SVZ spheres, but significantly lower in CB spheres. In addition, CB spheres stained weakly for specific epithelial markers. Ultrastructurally, CB spheres contained central immature unpolarized cells, and peripheral polarized more metabolic active cells. Immature adherence complexes were found both in both CB - and SVZ spheres, while desmosomal-like and tight junctions-like structures only were found in CB spheres.
Cultivation of adult human CB - and SVZ spheres creates a characteristic in vitro stem cell niche. CB spheres constite both neural - and epithelial characteristics, and the progenitors are connected through different types of immature junction-like structures.
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