Figure 4

The effects of different dwelling times on cell morphology and cytoskeleton organization. A) (a & e) On day 3, the mesothelial cells (MCs) appeared to be epithelioid-like in the phosphate-buffered saline (PBS) group and fibroblast-like in the transforming growth factor-β1 (TGF-β1) group. (b, c, d) There were some fibroblast-like MCs in the 8, 16, and 24 hour groups, and there were no significant differences between them. (g, h, i) Fibroblast-like MCs progressively increased after incubation with conventional dialysate for 6 days, especially in the case of the 24 hour group, where most of the MCs became fibroblast-like. Meanwhile, the cell count decreased with longer daily dwelling times, especially in the case of the 24 hour group (×200; N = 3). B) MC cytoskeletons were stained with anti-F-actin antibodies on days 3 and 6. (a & e) On day 3, the actin cytoskeleton in the cortical band of the PBS group was compared to the fibroblast-like cytoskeleton of the TGF-β1 group. (b, c, d) The intracellular F-actin distribution of the 8, 16, and 24 hour groups ranged between the morphology of the PBS and TGF-β1 groups. However, there were no significant differences between these three groups. (f & k) On day 6, the actin cytoskeleton in the cortical band of the PBS group was compared to the fibroblast-like cytoskeleton of the TGF-β1 group. (g & h) The intracellular F-actin distributions of the 8 and 16 hour groups were similar to those of the PBS group. (i & j) However, most MCs shrunk in the 24 hour group, and the remainder of the MCs had a fibroblast-like cytoskeleton similar to that exhibited in the TGF-β1 group (N = 3).