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Senescence in cultured trabecular meshwork cells
  1. Yukari Yamazaki,
  2. Hiroshi Matsunaga,
  3. Maki Nishikawa,
  4. Akira Ando (andoak{at}takii.kmu.ac.jp),
  5. Shiho Kaneko,
  6. Koji Okuda,
  7. Mitsumasa Wada,
  8. Seiji Ito,
  9. Miyo Matsumura
  1. Kansai Medical University, Japan
  2. Kansai Medical University, Japan
  3. Kansai Medical University, Japan
  4. Kansai Medical University, Japan
  5. Kansai Medical University, Japan
  6. Kansai Medical University, Japan
  7. Kansai Medical University, Japan
  8. Kansai Medical University, Japan
  9. Kansai Medical University, Japan

    Abstract

    Background/aim: It has been suggested that replicative senescence might be involved in the pathophysiology of age related diseases. We studied the process of senescence in trabecular meshwork (TM) cells.

    Methods: Porcine TM tissues were obtained and placed in primary cultures with DMEM/F12 medium. After 2 to 3 weeks, migrated and proliferated TM cells were trypsinized and cultured in serial passages, and identified with fluorescein-labeled low-density lipoprotein (DiI-Ac-LDL), a marker of TM cells. Staining for senescence-related β-galactosidase activity was performed at population doubling level (PDL) 2, 8, and 16 at pH 6. Terminal restriction fragment (TRF) length was examined by Southern blot analysis using a 32P-labeled telomere-specific sequence (TTAGGG)3 at each PDL.

    Results: DiI-Ac-LDL staining revealed that most (nearly 100%) of the cells in the culture were TM cells, which were flattened in shape and positive for senescence-related β-galactosidase staining at PDL 16. Reduction of TRF length as a function of population doubling was also shown.

    Conclusions: TM cells exhibited characteristics of senescence at PDL 16 in vitro. Our results demonstrated that cellular senescence may be related to the pathophysiology of primary open angle glaucoma.

    • beta-galactosidase
    • cellular senescence
    • primary open angle glaucoma
    • trabecular meshwork cell

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