Monitoring of morphonuclear characteristics of hormone-sensitive and insensitive HCT-15 and LOVO human colorectal cells by means of digital cell image analysis.
We recently set up in vitro several human colorectal neoplastic cell lines that we labelled hormone-sensitive (HS) in comparison with the original cell lines which appeared to be rather "hormone-insensitive" (HI). We describe here the cell proliferation rate and the morphonuclear characteristics of the HS and HI variants of the HCT-15 and LOVO human neoplastic colorectal cell lines which were cultured either in serum-supplemented or chemically-defined media. Morphonuclear characteristics were monitored by means of the digital cell image analyses of Feulgen-stained nuclei, while the proliferation activity of the various cell types was assessed by means of the tetrazolium-based compound (MTT) assay. The results show that it is possible to culture human LOVO and HCT-15 colorectal cells in chemically defined media. This said, growth in a chemically defined medium is difficult for these cells and is markedly less sustained than when they are cultured in serum-supplemented media. The transition of the culture from a serum-supplemented medium to a chemically defined one is accompanied by a very marked drop in cell proliferation and a number of profound changes in terms of morphonuclear characteristics. These changes basically involve chromatin decondensation, which occurs as the result of the drop in the number of large chromatin clumps in the nucleus.
Darro
F
F
Division de Cancérologie Expérimentale 1, Centre de Recherche Pierre Fabre, Castres, France.
Kruczynski
A
A
Etievant
C
C
Martinez
J
J
Kiss
R
R
D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus; D015179:Colorectal Neoplasms / Q000473:pathology*; D004396:Coloring Agents; D003470:Culture Media; D004958:Estradiol / Q000494:pharmacology; D005755:Gastrins / Q000494:pharmacology; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D009376:Neoplasms, Hormone-Dependent / Q000473:pathology*; D012394:Rosaniline Dyes; D013194:Staining and Labeling / Q000379:methods; D013268:Stimulation, Chemical; D013778:Tetrazolium Salts; D013844:Thiazoles; D014407:Tumor Cells, Cultured
D004396:Coloring Agents; D003470:Culture Media; C007722:Feulgen stain; D005755:Gastrins; D012394:Rosaniline Dyes; D013778:Tetrazolium Salts; D013844:Thiazoles; D004958:Estradiol; C022616:thiazolyl blue