Open AccessDifferent quiescence states of three culture cell lines detected by acridine orange staining of cellular RNA
Author(s) -
Baisch Heinz
Publication year - 1988
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990090409
Subject(s) - acridine orange , microbiology and biotechnology , rna , staining , trypan blue , hemocytometer , cell culture , biology , chinese hamster ovary cell , cell cycle , cell division , plating efficiency , cell , cell growth , biochemistry , genetics , gene
Three cell lines (CHO, L‐929 and R1H) were investigated for their growth kinetics and the difference of exponential and quiescent state of monolayers in medium with and without serum (L‐929). The noncycling populations of L‐929 and R1H in medium with serum contained increased G 1 ‐phase percentages but also considerable proportions of S Q and G 2Q cells. Although about 90% of the cells excluded trypan blue, the viability tested by colony assay was clearly lower than for exponentially growing cultures. CHO cells showed similar fractions of cells in G 1 ‐, S‐, and G 2 ‐Q compartments but in addition considerable cell loss. The RNA content of these cells was reduced in plateau phase by 7–48% depending on cell type and residence time in the noncycling state. The data suggest that the cells suffered from nutrition depletion and were arrested in all phases of the cycle. In contrast, L‐929 cells in medium without serum reduced their RNA content down to one‐third that of proliferating cells and still retained the full viability as shown by the same plating efficiency in a colony assay. Since about 90% of the cells had G 1 DNA content, these cells resemble true G 1 Q or G 0 cells controlled by growth factors rather than nutritional depletion.