Fig. 4.

Primary human keratocyte cultures in ERI medium supplemented with F-AME or C-AME. a Phase contrast micrographs showing CSK morphology after 5 passages in culture. Primary CSKs were prepared using donor corneal stromal tissue, HC778 and HC787. b Cell viability by calcein AM staining and cell quantification. Data are presented as mean and standard deviation for 4 primary CSK cultures. c Cell proliferation by EdU incorporation assay and cell quantification (n = 4 primary CSK cultures). d Wide-field spinning disk confocal laser microscopy for CSKs immunostained for ALDH3A1 (green fluorescence) and CD34 (red fluorescence). e Cell quantitation for the percentage of CSKs expressing ALDH3A1 and CD34 by confocal immunofluorescence in 4 primary CSK cultures. f Mean percentages of CSKs expressing ALDH3A1 and CD34 by immunostaining. Error bars: standard deviation. Scale bars: 300 μm (a), 500 μm (d)