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Sharada Paudel Kattel, Sowmya Parameswaran, Iqbal Ahmad; Molecular and Functional comparison of limbal iPS cell- and retina-derived RGCs. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1380.
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© ARVO (1962-2015); The Authors (2016-present)
The induced pluripotent stem (iPS) cells represent a viable source of retinal progenitors for regenerative medicine for the retina (Parameswaran et al., 2010, stem cells, 28:695-703). To address the barrier of insertional mutagenesis associated with nuclieic-acid-derived iPS cells, we reprogrammed limbal cells to pluripotency by non-nucleic acid means (Balasubramanian et al., 2009, Stem Cells, 27:3053-3062), which is a robust source of retinal progenitors for generating RGCs. The therapeutic utility of limbal iPS cell-derived RGCs will depend upon its acquisition of properties of RGCs present in the retina. Here we have compared the properties of limbal iPS cell-derived RGCs with that of RGCs present in the adult rat retina.
Rat limbal iPS cells generation and their subsequent neural differentiation were achieved as previously described (Balasubramanian et al., 2009). Limbal iPS cell-derived retinal progenitors were cultured in the presence of rat E14 retinal cell conditioned medium (E14CM) to generate RGCs. RGCs from the adult retina were obtained by the standard magnetic bead enrichment method using Thy1 antibody. The two RGC populations were subjected to immunocytochemical, Q-PCR and microarray analysis. In addition, to evaluate their target specificity they were co-cultured with superior/inferior colliculus (SC/IC) explants.
The limbal iPS cell-derived RGCs, like their retinal counterparts expressed regulators and markers of RGCs. In addition, they expressed molecules necessary for axonal pathfinding. However, their morphology was not as elaborate as RGCs enriched from the adult retina. Their processes like that of RGCs from the adult retina, could discriminate between SC and IC cells in establishing contacts. Preliminary comparative microarray analyses revealed reprogramming of the genome that approximated that of RGC obtained from the adult retina.
Our preliminary comparative analysis suggests that limbal iPS cell-derived RGCs embodies properties similar to that of RGCs derived from the adult retina and therefore may also function similarly.
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