title: Decellularized retinal matrix: Natural platforms for human retinal progenitor cell culture.
abstract: Tissue decellularization strategies have enabled engineering of scaffolds that preserve native extracellular matrix (ECM) structure and composition. In this study, we developed decellularized retina (decell-retina) thin films. We hypothesized that these films, mimicking the retina niche, would promote human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. Retinas isolated from bovine eyes were decellularized using 1% w/v sodium dodecyl sulfate (SDS) and pepsin digested. The resulting decell-retina was biochemically assayed for composition and cast dried to develop thin films. Attachment, viability, morphology, proliferation and gene expression of hRPC cultured on the films were studied in vitro. Biochemical analyses of decell-retina compared to native retina indicated the bulk of DNA (94%) was removed, while the majority of sulfated GAGs (55%), collagen (83%), hyaluronic acid (87%), and key growth factors were retained. The decell-retina films supported hRPC attachment and growth, with cell number increasing 1.5-fold over a week. RT-PCR analysis revealed hRPC expression of rhodopsin, rod outer membrane, neural retina-specific leucine zipper neural and cone-rod homeobox gene on decell-retina films, indicating photoreceptor development. In conclusion, novel decell-retina films show promise as potential substrates for culture and/or transplantation of retinal progenitor cells to treat retinal degenerative disorders.
  
  
  CONCLUSIONS
  In this study, we report the development of a novel biomaterial, based on decellularized retina (decell-retina) that mimics the retina niche and promotes human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. We estimated, for the first time, the amounts of collagen I, GAGs and HA present in native retina, as well as the decell-retina. We demonstrated that retinas can be decellularized using ionic detergents and can be processed into mechanically stable thin films, which can act as substrates for culturing hRPCs. Rhodopsin, ROM1, NRL and CRX gene expression on the decell-retina films indicated photoreceptor development from RPCs. These results support the potential of decell-retina as a cell delivery platform to treat and manage retinal degenerative disease like AMD.
authors.affiliation: Department of Chemical Engineering, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA; The Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA. Electronic address: j.kundu@neu.edu.
authors.forename: Joydip
authors.initials: J
authors.lastname: Kundu
authors.affiliation: Department of Chemical Engineering, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA. Electronic address: malachad@gmail.com.
authors.forename: Andrew
authors.initials: A
authors.lastname: Michaelson
authors.affiliation: Department of Chemical Engineering, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA. Electronic address: talbot.kristen@gmail.com.
authors.forename: Kristen
authors.initials: K
authors.lastname: Talbot
authors.affiliation: The Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA. Electronic address: petr.baranov@schepens.harvard.edu.
authors.forename: Petr
authors.initials: P
authors.lastname: Baranov
authors.affiliation: The Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA. Electronic address: michael.young@schepens.harvard.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Young
authors.affiliation: Department of Chemical Engineering, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA. Electronic address: rebecca@coe.neu.edu.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Carrier
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002417:Cattle; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D004247:DNA / Q000737:chemistry; D005109:Extracellular Matrix / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D016350:Leucine Zippers; D008855:Microscopy, Electron, Scanning; D008858:Microscopy, Phase-Contrast; D009474:Neurons / Q000378:metabolism; D010434:Pepsin A / Q000737:chemistry; D012160:Retina / Q000166:cytology* / Q000378:metabolism*; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism; D012243:Rhodopsin / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D012967:Sodium Dodecyl Sulfate; D004247:DNA; D012243:Rhodopsin; D010434:Pepsin A
keywords: Decellularization; Extracellular matrix (ECM); Retina; Retinal progenitor cell (RPC); Tissue engineering