Figure 6. Human CE cells form a high density monolayer and express ZO-1 when cultured on a COL4 EBM.

(a) In contrast, human CE cells isolated from two corneas from two separate donors were cultured for >25 days on two COL4+LAM EBMs (1 donor pair of corneas per EBM) and in each case failed to become for a full confluent monolayer, expressed no ZO-1 at the cell borders, and the F-actin cytoskeleton of the cells has large stress fibers spanning across the cell bodies. The LAM was antibody labeled and shows the structure of the LAM does not match that which was initially present in the EBM or what was produced by the bovine CE cells. (b) Human CE cells isolated from single corneas from two separate donors were cultured for 14 days on two COL4 EBMs (1 donor per EBM) and formed monolayers with a densities of 1700 cells mm⁻² (top row) and 760 cells mm⁻². The cells express continuous ZO-1 at the cell borders. The F-actin cytoskeleton shows very organized stress fibers located cortically. In most regions of the EBM the COL4 (labeled during fabrication) was gone indicating the cells had completely remodeled the COL4, however it did remain in some areas. Scale bars: 50μm.