Figure 2
Evaluation of clonogenic potential and cell morphology in prolonged airway epithelial cell cultures. (a) Representative images of CFE assay used for the determination of clonogenic and aborted colonies. (b) Quantification of the percentage of clonogenic cells (grown colonies, left) and aborted colonies (terminally differentiated colonies, right) respectively out of the total cells seeded and out of the total grown colonies in the indicator dish. No significant differences were observed in clonogenic cells (human vs porcine trachea p-value = 0.88; human vs porcine bronchus p-value = 0.33) and aborted colonies (human vs porcine trachea p-value = 0.27; human vs porcine bronchus p-value = 0.20). (c) Representative images of human and porcine tracheal epithelial cell cultures at starting (pII) and final (pIX) passages. (d) CFE assay evaluates the number of clonogenic and aborted colonies at starting and final sub-cultures. (e) Quantification of the grown (left) and aborted (right) colonies in the CFE dishes. No significant differences were observed in the clonogenic potential of human and porcine cells in second (human vs porcine trachea p-value = 0.71; human vs porcine bronchus p-value = 0.60) and ninth expansion passage (human vs porcine trachea p-value = 0.98; human vs porcine bronchus p-value = 0.68). Similarly, no significant differences were observed in the aborted colonies percentage in second (human vs porcine trachea p-value = 0.25; human vs porcine bronchus p-value = 0.25) and ninth expansion passage (human vs porcine trachea p-value = 0.07; human vs porcine bronchus p-value = 0.10). The results are shown as means ± SD of six human samples (n = 3 trachea and n = 3 bronchi) and four porcine samples (n = 2 trachea and n = 2 bronchi).
