title: Fabrication of degradable polymer scaffolds to direct the integration and differentiation of retinal progenitors.
abstract: Retinal progenitor cells (RPCs) are self-renewing cells capable of differentiating into the different retinal cell types including photoreceptors, and they have shown promise as a source of replacement cells in experimental models of retinal degeneration. We hypothesized that a biodegradable polymer scaffold could deliver these cells to the subretinal space in a more organized manner than bolus injections, while also providing the graft with laminar organization and structural guidance channels. We fabricated highly porous scaffolds from blends of poly(L-lactic acid) and poly(lactic-co-glycolic acid) using a variety of techniques to produce pores oriented normal to the plane of the scaffold. RPCs were seeded on the polymer scaffolds and cultured for 14 days. Seeded scaffolds were then either fixed for characterization or used in an explant or in vivo rat model. The scaffolds were fully covered by RPCs in 3 days. Attachment of RPCs to the polymer scaffold was associated with down-regulation of immature markers and up-regulation of markers of differentiation. This suggests that the scaffold may promote differentiation of RPCs. The seeded cells elaborated cellular processes and aligned in the scaffold in conjunction with degenerating retinal explants. The cells also exhibited morphologies consistent with photoreceptors including a high degree of polarization of the cells. This data suggests that the scaffold may be a means to assist in the promotion of photoreceptor phenotypes. Implantation of the seeded scaffold into the rat eye is associated with increased RPC survival. Taken together, these data suggest that these polymer scaffolds provide a useful means for delivering RPCs to the subretinal space and may assist in the formation of retinal cell phenotypes, although it is clear that more cues are needed to direct the differentiation of RPCs into functional photoreceptors.
authors.affiliation: Biomedical Engineering, Yale University, 15 Prospect Street, Becton Center 229, New Haven, CT 06511, USA. erin.lavik@yale.edu
authors.forename: E B
authors.initials: EB
authors.lastname: Lavik
authors.forename: H
authors.initials: H
authors.lastname: Klassen
authors.forename: K
authors.initials: K
authors.lastname: Warfvinge
authors.forename: R
authors.initials: R
authors.lastname: Langer
authors.forename: M J
authors.initials: MJ
authors.lastname: Young
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002454:Cell Differentiation; D017690:Cell Transplantation; D018920:Coculture Techniques; D015536:Down-Regulation; D048091:Guided Tissue Regeneration / Q000379:methods; D007150:Immunohistochemistry; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D010641:Phenotype; D010786:Photoreceptor Cells / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000166:cytology* / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D001672:Biocompatible Materials; D011108:Polymers