OBJECTIVE: To observe changes of the cytoskeleton of cultured human retinal pigment epithelium (hRPE) cells in a mechanical stress model in vitro. METHODS: Ferric oxide beads, coated with collagen, were added to dishes containing substrate-attached hRPE cells. After incubation, the cells were washed to remove unbound beads. The cells were pretreated by actin polymerization inhibitor cytochalasin D (0.05 mmol/L, 25 min). Before and 4, 8, 12 and 24 h after vertical magnetic force application the cells were stained for actin and vimentin using immunity fluorescence double labelled technique and analyzed with confocal microscopy. RESULTS: Four hours after force application, the polarity of hRPE cells was changed, and actin was polymerized into filaments in array of them along with the direction of the force. Cytoskeleton filaments were located mainly around the nucleus and the ferric oxide beads. Pretreatment with cytochalasin D, the same changes as described above occurred 8 h after magnetic force application. CONCLUSIONS: Mechanical force could induce changes of distribution of cytoskeleton in hRPE cells, which are some biological characteristics of muscle cells, and this may facilitate the cellular migration and proliferation.
OBJECTIVE: To observe changes of the cytoskeleton of cultured human retinal pigment epithelium (hRPE) cells in a mechanical stress model in vitro. METHODS:Ferric oxide beads, coated with collagen, were added to dishes containing substrate-attached hRPE cells. After incubation, the cells were washed to remove unbound beads. The cells were pretreated by actin polymerization inhibitor cytochalasin D (0.05 mmol/L, 25 min). Before and 4, 8, 12 and 24 h after vertical magnetic force application the cells were stained for actin and vimentin using immunity fluorescence double labelled technique and analyzed with confocal microscopy. RESULTS: Four hours after force application, the polarity of hRPE cells was changed, and actin was polymerized into filaments in array of them along with the direction of the force. Cytoskeleton filaments were located mainly around the nucleus and the ferric oxide beads. Pretreatment with cytochalasin D, the same changes as described above occurred 8 h after magnetic force application. CONCLUSIONS: Mechanical force could induce changes of distribution of cytoskeleton in hRPE cells, which are some biological characteristics of muscle cells, and this may facilitate the cellular migration and proliferation.