Objective: To characterize the Integrin-α6β1 expressing clone for its proliferative, clonogenic capabilities and immunohistochemical marker expression.
Methods: rats were subjected to short heat stress followed by isolation of salivary gland parenchyme using the Percoll gradient. Thereafter, Integrin-α6β1 positive cells were secluded by MACS.Those cells were cultivated and clones isolation was done by limiting dilution. To examine the clonogenic capability of the cells, clones were counter-stained and counted. specific cell markers were detected by immunohistochemistry using antibodies. To assess tumorogenic proliferation of the clone, the growth in soft agar was compared with cell line.
Results: Integrin-α6β1 positive cells had been from doesens of animals in separate preparations. Only one preparation survived passage 24 and cloned by limiting dilution (RSC-rat salivary clone). The clone obtained found to divide rapidly and do not undergo senescence in high passages. The clonogenic capability of RSC was 5-10 times more compared to normal cells. Immunohistochemical stain of RSC detected the presence of Integrin-α6β1 while P63 was missing, unlike control cells which express both markers. Soft agar test has shown that RSC grow slowly on soft agar.
Conclusions:
RSC clonogenic capability and divisions potential was found higher than “normal” cells.
RSC express á6â1 integrins indicating basal origin, but do not express P-63, which is a stem cells marker.
RSC grow on soft agar, unlike normal epithelial cell indicating being immortal.