In Vitro Cellular & Developmental Biology

, Volume 21, Issue 5, pp 282–287 | Cite as

Analysis of the cytotoxic effects of light-exposed hepes-containing culture medium

  • J. S. ZiglerJr.
  • J. L. Lepe-Zuniga
  • B. Vistica
  • I. Gery


The addition ofN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES) to RPMI 1640 medium markedly increases the production of cytotoxic products during exposure of the medium to visible light. The cytotoxicity has been analyzed by measuring uptake of [3H]thymidine by murine thymocytes cultured in preirradiated medium containing 25 mM HEPES. Complete inhibition of thymidine uptake was produced by exposing 50% of the culture medium to light for 3 h before addition of cells. The HEPES-mediated effect requires only that HEPES and riboflavin be exposed to light; other medium constituents are not necessary. Hydrogen peroxide is a principal cytotoxic agent produced in this system. It is demonstrated that most, but not all, of the inhibition of thymidine uptake can be attributed to hydrogen peroxide.

Key words

tissue culture medium HEPES light-induced cytotoxicity hydrogen peroxide 


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Copyright information

© Tissue Culture Association, Inc 1985

Authors and Affiliations

  • J. S. ZiglerJr.
    • 1
  • J. L. Lepe-Zuniga
    • 1
  • B. Vistica
    • 1
  • I. Gery
    • 1
  1. 1.Laboratory of Vision ResearchNational Eye InstituteBethesda

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