Figure 1. Morphological, histological and ultrastructural characterization of primary cultured GCD2 corneal fibroblasts. (A) Phase-contrast micrograph of WT and GCD2 HO corneal fibroblasts. Cells were analyzed and photographed using an inverted microscope. The GCD2 HO corneal fibroblasts showed a number of morphological abnormalities that are different from WT corneal fibroblasts, and the GCD2 cells contain different-sized intracellular deposits. The increased granularity within the cells was not seen in WT cells. (B) WT and GCD2 HO corneal fibroblasts were costained with anti-TGFBI and anti-CTSD antibodies. GCD2 homozygous corneal fibroblasts showed much more extensive pattern of colocalized mut-TGFBI with CTSD than observed in WT corneal fibroblasts. (C) Electron microscopy (EM) of WT and GCD corneal fibroblasts. Cells were fixed and processed for EM. Many abnormal mitochondria (short or fragmented) were observed in GCD2 cells compared with WT cells. Bars: 2000 nm. (D) Quantitation of mitochondrial numbers. Mitochondrial numbers were measured in six WT cells and six GCD2 cells. Results represent the mean ± SD *p < 0.05; **p < 0.01.