Cytofluorometric nuclear DNA determinations on human corneal endothelial cells.

In order to investigate polyploidization of human corneal endothelium, nuclear DNA cytofluorometry was performed. Touch smears of 31 human corneal endothelium taken from 21 autopsies and 10 donor eyes were fixed with absolute methanol and subsequently stained by the pararosaniline-Feulgen nuclear reaction. The optimum condition for Feulgen hydrolysis of the specimens was determined, and all smears were hydrolysed in 1 N HCl for 5 min at 60 degrees C. After the Feulgen reaction, the pararosaniline-DNA fluorescence was measured with an epi-illumination cytofluorometer (Olympus AH-QRFL). Polyploid nuclei, which are the result of abortive cell division after DNA synthesis, appeared with DNA contents of tetraploid (19 cases) and octaploid (two cases) classes. All other nuclei had DNA quantities within the range of the diploid class. Nuclei containing DNA between diploid and tetraploid amounts were not found. It was concluded that most human corneal endothelial cells are of post mitotic Gl population but that a few are in the generative G0 phase.