Title of Oral/Poster Presentation

Characterizing the Growth of Retinal Pigment Epithelial Cells on Recombinant Spider Silk Membranes

Class

Article

College

College of Engineering

Faculty Mentor

Elizabeth Vargis

Presentation Type

Poster Presentation

Abstract

Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. Major participants in AMD are the retinal pigment epithelial (RPE) cells. They grow posterior to the photoreceptors on an acellular protein structure, called Bruch’s membrane. These cells transport nutrients, remove waste, and provide support for the photoreceptor cells. As ageing occurs, a lipid substance called drusen forms inside Bruch’s membrane and inhibits RPE cell growth and function. To better understand the relationship between the disruption of RPE cells and AMD, cells must be grown and characterized in vitro. Such research could lead to the development of an in vitro model of the eye, which would be used to research potential diagnostic methods, treatments, or medications to counteract the effects of AMD. However, this model requires the use of a synthetic Bruch’s membrane that also can mimic natural structure and function of the retina. A material for the formation of a proteinaceous film or membrane was developed from recombinant spider silk proteins (rSSps) and used as a substrate for RPE growth. Free-standing, semipermeable films were engineered using multiple techniques to ensure a rSSp membrane that is biomimetic to Bruch’s membrane. ARPE-19 cell proliferation was analyzed using a PicoGreen double stranded DNA assay. Immunocytochemistry staining and scanning electron microscopy (SEM) were employed to assess the morphology of cells and a permeability assay was used to evaluate the formation of tight junctions. After analysis, it was found that the free-standing rSSps membranes supported the growth of the immortalized ARPE-19 cells. Based upon these results, the application of rSSps may prove beneficial for in vitro modeling of Bruch’s membrane and AMD. Ongoing work will focus on using primary porcine RPE cells to create a more realistic retina model as they produce structures, like microvilli, that are not expressed in ARPE-19 cells.

Location

The North Atrium

Start Date

4-12-2018 3:00 PM

End Date

4-12-2018 4:15 PM

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Apr 12th, 3:00 PM Apr 12th, 4:15 PM

Characterizing the Growth of Retinal Pigment Epithelial Cells on Recombinant Spider Silk Membranes

The North Atrium

Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. Major participants in AMD are the retinal pigment epithelial (RPE) cells. They grow posterior to the photoreceptors on an acellular protein structure, called Bruch’s membrane. These cells transport nutrients, remove waste, and provide support for the photoreceptor cells. As ageing occurs, a lipid substance called drusen forms inside Bruch’s membrane and inhibits RPE cell growth and function. To better understand the relationship between the disruption of RPE cells and AMD, cells must be grown and characterized in vitro. Such research could lead to the development of an in vitro model of the eye, which would be used to research potential diagnostic methods, treatments, or medications to counteract the effects of AMD. However, this model requires the use of a synthetic Bruch’s membrane that also can mimic natural structure and function of the retina. A material for the formation of a proteinaceous film or membrane was developed from recombinant spider silk proteins (rSSps) and used as a substrate for RPE growth. Free-standing, semipermeable films were engineered using multiple techniques to ensure a rSSp membrane that is biomimetic to Bruch’s membrane. ARPE-19 cell proliferation was analyzed using a PicoGreen double stranded DNA assay. Immunocytochemistry staining and scanning electron microscopy (SEM) were employed to assess the morphology of cells and a permeability assay was used to evaluate the formation of tight junctions. After analysis, it was found that the free-standing rSSps membranes supported the growth of the immortalized ARPE-19 cells. Based upon these results, the application of rSSps may prove beneficial for in vitro modeling of Bruch’s membrane and AMD. Ongoing work will focus on using primary porcine RPE cells to create a more realistic retina model as they produce structures, like microvilli, that are not expressed in ARPE-19 cells.