May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
An In Vitro Study of the effect of Varying pH on survival of Retinal Ganglion Cells
Author Affiliations & Notes
  • W.W. Phillips
    Ophthalmology, University of Florida College of Medicine, Jacksonville, FL
  • V.A. Shah
    Ophthalmology, University of Florida College of Medicine, Jacksonville, FL
  • B. Tripathi
    Ophthalmology, University of South Carolina School of Medicine, Columbia, SC
  • R. Tripathi
    Ophthalmology, University of South Carolina School of Medicine, Columbia, SC
  • K.V. Chalam
    Ophthalmology, University of Florida College of Medicine, Jacksonville, FL
  • Footnotes
    Commercial Relationships  W.W. Phillips, None; V.A. Shah, None; B. Tripathi, None; R. Tripathi, None; K.V. Chalam, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 875. doi:
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      W.W. Phillips, V.A. Shah, B. Tripathi, R. Tripathi, K.V. Chalam; An In Vitro Study of the effect of Varying pH on survival of Retinal Ganglion Cells . Invest. Ophthalmol. Vis. Sci. 2004;45(13):875.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: To evaluate retinal ganglion cell (RGC) viability at various pH concentrations and time intervals. Methods: RGCs cultured using Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum and 1% streptomycin/penicillin were allowed to grow to confluence at 37°C for 3 days. To maintain a continuous pH for cellular growth, 5% CO2 was provided for incubation. The pH concentrations of the cell–lines media was adjusted to values of 7.0 and 7.8; then allowed to incubate for 2, 4, and 6 hours. Cellular viability was assessed using Fluorescein Isothiocyanate (FITC) labeled Annexin V and Propidium iodide by flow cytometry (Becton Dickinson, Mountain View, CA). Cellular integrity in cultured RGCs was evaluated by flow cytometric analysis. Results: Results were expressed as positive or negative based on a shift in peak channel fluorescence intensity of the experimental cultures with respect to control. Apoptotic detection was also confirmed morphologically by light–microscopy . Flow cytometric analysis of Annexin–V for control and experimental cultures are listed in table 1.Exposure to various pH concentrations at specific times altered membrane activity (p<0.01). In addition flow cytometry detected a decrease in forward scatter, increase in side scatter and propidium iodide fluorescence, all physical characteristics of apoptosis. Conclusion: Increase in pH in cultured media induces cell–death in ganglion cells and is exponentially related to degree of increase in pH. Table 1 

Keywords: apoptosis/cell death • ganglion cells 
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