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Caio V. Regatieri, Petr Baranov, Jing Yao, Chi Wan Ko, Sarah Tao, Michael J. Young; Biodegradable Polycaprolactone Scaffolds with Integrated Microtopography Promote Differentiation of Human Retinal Progenitor Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5896.
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© ARVO (1962-2015); The Authors (2016-present)
Retinal degenerations cause permanent visual loss and affect millions of people worldwide. Previous work has demonstrated the utility of using biodegradable polymer scaffolds to induce differentiation and deliver retinal progenitor cells for cell replacement therapy. In this study, we engineered a biodegradable polycaprolactone (PCL) thin film scaffold with integrated sub-micron topography and analyzed its effect on the morphology and differentiation of human retinal progenitor cells (hRPCs).
The polymer was microfabricated with ridge-grooves or posts on the surface. hRPCs were isolated from human retina of 14 to 18 weeks gestational age and expanded in vitro, in low-tension oxygen (3%). At passage five the cells were seeded in an 8-well slide (control group) and on three different types of PCL scaffolds (smooth surface, ridge-groove and post). After 24 hours of culture, the cell morphology was evaluated. After one-week, real time polymerase chain reaction (PCR) and immunocytochemistry (ICC) assays were performed on hRPCs cultured on the biodegradable polymer and on the control group, in order to evaluate the differentiation of hRPCs into photoreceptors.
Microfabricated topography in a PCL thin film enhanced the attachment and organization of hRPCs compared to unstructured surfaces. After 24 hours of culture hRPCs cultured on a ridge-groove topography developed substantial elongation and parallel alignment. No morphological changes were observed in the other groups. hRPCs adherent to PCL differentiated toward mature photoreceptor phenotypes as evidenced by changes in mRNA and protein levels. Using real time quantitative PCR and ICC we observed a statistically significant upregulation in the expression of rhodopsin, CRX, recoverin and a statistically significant downregulation in SOX2 (a marker for undifferentiated progenitor cells) and PAX6 compared to cells grown on polystyrene.
This unique structured PCL thin-film scaffold provides a means to organize and differentiate RPCs in a controlled manner.Copyright © 2010 by The Charles Stark Draper Laboratory, Inc and Schepens Eye Research Institute all rights reserved.
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