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Cell culture conditions affect RPE phagocytic function.

Mike O Karl1, Monika Valtink, Jürgen Bednarz

  • 1Department of Ophthalmology, Cornea Bank, University Eye Clinic Eppendorf, Martinistrasse 52, Hamburg 20246, Germany. mail@mikeology.de

Graefe'S Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Fur Klinische Und Experimentelle Ophthalmologie
|December 21, 2006
PubMed
Summary

Serum-free media like hSFM and SFM enhance retinal pigment epithelium (RPE) cell phagocytosis. Supplements such as fetal calf serum (FCS) and retinal extract (RE) also improve RPE function in cell culture.

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Area of Science:

  • Ophthalmology
  • Cell Biology
  • Tissue Engineering

Background:

  • Retinal pigment epithelium (RPE) cell phenotype and function are influenced by culture conditions.
  • RPE tissue engineering aims for serum-free propagation of differentiated, functional RPE cells in vitro.

Purpose of the Study:

  • To compare the effects of various media and supplements on RPE cell phagocytosis.
  • To identify optimal conditions for maintaining RPE function in vitro for tissue engineering applications.

Main Methods:

  • Developed and utilized an optimized high-throughput phagocytosis assay.
  • Cultured adult human SV40-RPE cells in various serum-free media (MEM(E), DMEM, F99, SFM, hSFM) with or without supplements.
  • Measured phagocytic binding and uptake of SNAFL-2 labeled outer segments (OS).

Main Results:

  • Serum-free media hSFM and SFM promoted higher RPE phagocytosis compared to DMEM, MEM, and F99.
  • Choroid-conditioned medium (ChCM) dose-dependently decreased phagocytosis, while 1% retinal extract (RE) increased it.
  • Fetal calf serum (FCS) increased phagocytosis; however, 15% ChCM inhibited this effect, which was reversed by 1% RE.

Conclusions:

  • Cell culture media and environmental factors significantly alter RPE phagocytic ability.
  • Serum-free media SFM and hSFM were most effective in promoting phagocytosis.
  • FCS and 1% RE support RPE phagocytosis, while factors from retina and choroid have differential effects, potentially impacting retinal pathogenesis.