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.
- cellular senescence
- primary open angle glaucoma
- trabecular meshwork cell