December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Effects of Components of Serum on Apoptosis in Cultured Conjunctival Epithelial Cells
Author Affiliations & Notes
  • A Higuchi
    Ophthalmology
    Tokyo Dental College Ichikawa Japan
  • M Suematsu
    Biochemistry Keio University Tokyo Japan
  • H Aburatani
    Genome Science 3Research Center for Advanced Science and Technology University of Tokyo Tokyo Japan
  • K Tsubota
    Department of Ophthalmology
    Tokyo Dental College Ichikawa Japan
  • Footnotes
    Commercial Relationships   A. Higuchi, None; M. Suematsu, None; H. Aburatani, None; K. Tsubota, None.
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 3171. doi:
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      A Higuchi, M Suematsu, H Aburatani, K Tsubota; Effects of Components of Serum on Apoptosis in Cultured Conjunctival Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3171.

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

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Abstract

Abstract: : Purpose: Conjunctival epithelial cell line, CCL20.2 (CCL) requires fetal bovine serum (FBS) in the medium to survive. If serum is removed from medium, the cells will die. To elucidate the survival mechanism of serum components in CCL, we added components of serum to medium and estimated effects of components on anti-apoptosis and survive. Methods: CCL was grown in Medium 199 with 10% FBS to 60-70% confluent, and medium was changed to Medium 199 with 10% human serum, without FBS, or without FBS and with components of serum. After 24 hours incubation, we harvested the cells respectively, and measured cellular viability. The cells were solubilized and cell lysate was centrifuged to measure caspase activities and DNA fragmentation in supernatant. Results: Serum deprivation induced decrease of cellular viability (from 94 to 56 %) and increase of activities of caspase (from 0.12 to 0.47 n mol/hr/mg protein) and DNA fragmentation (from 0.027 to 0.11). This apoptosis repressed by components of serum, such as growth factors (30-40 %), vitamin (20-70 %), and serum proteins (30-70 %), and these anti-apoptotic effects act synergically. Conclusion: This study suggested cell death of serum deprivation was apoptosis with activation of caspases, and this apoptosis was suppressed by serum components.

Keywords: 323 apoptosis/cell death • 365 conjunctiva 
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