Fig. 5

Endothelial growth in culture. Immediately after isolation, cell clusters are visible (green arrow), where individual cells are distinctly visible, but microtubules remain intact. 2 h after isolation 0 DIV, cell somas (blue arrow) can be seen attached to the bottom of the plate around the cell clusters. By 2 DIV, the first media change is completed, indicated by the red triangle icon. Endothelial-specific morphology is seen as early as 0 DIV characterized by cells growing in circular patterns extremely close to one another. At 1 DIV, cell debris from dying non-endothelial and other non-attached cells are visible (purple arrow). Each endothelial cluster grows and extends to combine with other clusters. Depending on plating concentration, cells reach confluency and can be frozen as early as 5 DIV. Morphology does not change between 0 DIV and 1 DIV, cells simply become longer with the same phenotype with increased culture time. Scale bars in all images: 200 μm