Quantitation of in vitro ciliated cell growth through image analysis

Ciliated cell cultures can be produced in outgrowths from explants of human respiratory epithelium. An image analysis technique was developed to quantify the percentage of active ciliated cells present in these cultures. The subtraction 2 by 2 of five successive video images of the cultures, followed by the addition of the resulting images, allowed the determination of the culture surface covered by ciliated cells. The percentage of this surface varied according to the region of the explant (27.7% in the outgrowth near the explant and 4.1% at the periphery of the outgrowth). High variations were observed within the same region of an outgrowth, as well as from one outgrowth to another. However, maximal differentiation was observed after 4 d of culture. The quantitation technique described in the present work might be useful for studying in vitro the respiratory epithelial injury and the subsequent repair processes.