Influence Of Pyruvate And Sodium Bicarbonate On The Induction Of Pigmentation In Arising Retinal Pigmented Epithelial-19 Cells (Arpe-19)

Valerie Hinsch

School Name

Governor's School for Science and Math

Grade Level

12th Grade

Presentation Topic

Biochemistry

Presentation Type

Mentored

Mentor

Mentor: Dr. Ablonczy; Department of Ophthalmology, Medical University of South Carolina

Abstract

The retinal pigment epithelium (RPE) is responsible for the metabolic maintenance of the photoreceptors in the eye. Dysfunction of this tissue can induce conditions like macular edema which are currently without clinical treatment, creating the need for accurate models of RPE for successful development of treatments. ARPE-­19 cells, the most widely used researched cell line, lack the differentiation required to accurately model in vivo properties of the tissue, especially pigmentation. Multiple studies to reestablish ARPE-­19 cell qualities have remained unsuccessful. However, a recent paper reported the desired repigmentation of the cells (Ahmado et al., 2010), due to the presence of high concentrations of pyruvate and sodium bicarbonate in the media. The goal of this research was to replicate these experiments while testing the development of barrier function and the secretion of vascular endothelial growth factor (VEGF), expressed by RPE to maintain the ocular vasculature. This aim was achieved by growing the cells in two different media (DMEM/F-12 and DMEM/Pyruvate) with the same amount of sodium bicarbonate (3.7 g/L) and different amounts of pyruvate (55 and 110 mg/L). Barrier function was tested through transepithelial resistance using an epithelial voltohmmeter and VEGF secretion was determined from collecting the apical and basal media once a week. VEGF could not be tested due to time constraints, but results indicate that there was no pigmentation and a reduction in barrier function in the presence of pyruvate. This research did not validate previous work, and therefore must be repeated.

Location

Owens 203

Start Date

4-16-2016 10:00 AM

COinS
 
Apr 16th, 10:00 AM

Influence Of Pyruvate And Sodium Bicarbonate On The Induction Of Pigmentation In Arising Retinal Pigmented Epithelial-19 Cells (Arpe-19)

Owens 203

The retinal pigment epithelium (RPE) is responsible for the metabolic maintenance of the photoreceptors in the eye. Dysfunction of this tissue can induce conditions like macular edema which are currently without clinical treatment, creating the need for accurate models of RPE for successful development of treatments. ARPE-­19 cells, the most widely used researched cell line, lack the differentiation required to accurately model in vivo properties of the tissue, especially pigmentation. Multiple studies to reestablish ARPE-­19 cell qualities have remained unsuccessful. However, a recent paper reported the desired repigmentation of the cells (Ahmado et al., 2010), due to the presence of high concentrations of pyruvate and sodium bicarbonate in the media. The goal of this research was to replicate these experiments while testing the development of barrier function and the secretion of vascular endothelial growth factor (VEGF), expressed by RPE to maintain the ocular vasculature. This aim was achieved by growing the cells in two different media (DMEM/F-12 and DMEM/Pyruvate) with the same amount of sodium bicarbonate (3.7 g/L) and different amounts of pyruvate (55 and 110 mg/L). Barrier function was tested through transepithelial resistance using an epithelial voltohmmeter and VEGF secretion was determined from collecting the apical and basal media once a week. VEGF could not be tested due to time constraints, but results indicate that there was no pigmentation and a reduction in barrier function in the presence of pyruvate. This research did not validate previous work, and therefore must be repeated.