May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
The Effect of Pigment Epithelium Derived Factor (PEDF) on Human Retinal Pigment Epithelial Cell Proliferation and Intracellular Growth Factor Synthesis
Author Affiliations & Notes
  • R. Lahiri
    Department of Ophthalmology and Visual Sciences, WK Kellogg Eye Center, University of Michigan, Ann Arbor, MI
  • P. Kothary
    Department of Ophthalmology and Visual Sciences, WK Kellogg Eye Center, University of Michigan, Ann Arbor, MI
  • M. Del Monte
    Department of Ophthalmology and Visual Sciences, WK Kellogg Eye Center, University of Michigan, Ann Arbor, MI
  • Footnotes
    Commercial Relationships  R. Lahiri, None; P. Kothary, None; M. Del Monte, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 639. doi:
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      R. Lahiri, P. Kothary, M. Del Monte; The Effect of Pigment Epithelium Derived Factor (PEDF) on Human Retinal Pigment Epithelial Cell Proliferation and Intracellular Growth Factor Synthesis . Invest. Ophthalmol. Vis. Sci. 2004;45(13):639.

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Abstract

Abstract: : Purpose: PEDF is a growth factor for human retinal pigment epithelial (hRPE) cells and has been implicated in the pathology of proliferative eye disease. The purpose of this study is to determine if PEDF inhibits fetal bovine serum (FBS) stimulated cultured hRPE cell proliferation and the synthesis of other intracellular growth factors, such as fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor (VEGF). Methods: Primary cultures of hRPE cells were established from four human eyes obtained from the Michigan Eye Bank. The proliferative activity of sub–confluent hRPE cells was determined by 3H–thymidine incorporation and the trypan blue exclusion method. The level of intracellular FGF2 and VEGF synthesis was monitored by immunocytochemistry using FGF2 and VEGF specific antibodies. Data were analyzed by students T test, and p<0.05 was indicative of a significant difference. Results: Measurement of hRPE cell proliferation by direct cell counting using the trypan blue exclusion method and by 3H–thymidine incorporation revealed increased proliferative activity in hRPE cells grown in the presence of fetal bovine serum (FBS) when compared with control (35,346 ± 5,415 cpm vs 26,798 ± 4,616 cpm, p<0.05, n=4) , while those exposed to increasing doses of PEDF in the presence of FBS showed decreased proliferative activity (27,217 ± 4186 cpm vs 35,346 ± 5415 cpm, p<0.05, n=4). Furthermore, immunocytochemistry revealed that PEDF decreased the amount of FBS–stimulated FGF2 and VEGF synthesis. Cells treated with FBS alone exhibited brighter and more widespread FGF2 and VEGF staining, while fluorescence was decreased compared to controls in cells treated with PEDF. Conclusions:1) PEDF inhibits FBS–stimulated hRPE cell proliferation. 2) PEDF decreases FBS–induced synthesis of FGF2 and VEGF. This data suggests that PEDF is a negative regulator of hRPE cell proliferation by decreasing the synthesis of intracellular growth factors. PEDF concentration in ocular fluid may play an important role in the pathogenesis of proliferative eye disease.

Keywords: retinal pigment epithelium • growth factors/growth factor receptors 
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