Global analysis of endothelial cell line proliferation patterns based on nutrient-depletion models: implications for a standardization of cell proliferation assays.
It is known that cell populations growing in different environmental conditions may exhibit different proliferation patterns. However, it is not clear if, despite the diversity of the so-observed patterns, inherent cellular growth characteristics of the population can nevertheless be determined. This study quantifies the proliferative behaviour of the permanent endothelial human cell line, Eahy926, and establishes to which extent the estimation of the cell proliferation rate depends on variations of the experimental protocols. Cell proliferation curves were obtained for cells cultured over 16 days and the influences of cell seeding densities, foetal bovine serum content and frequency of culture medium changes were investigated. Quantitative dynamic modelling was conducted to evaluate the kinetic characteristics of this cell population. We proposed successive models and retained a nutrient-depletion toxicity dependant model, which takes into account the progressive depletion of nutrients, as well as the increase of toxicity in the cell culture medium. This model is shown to provide a very good and robust prediction of the experimental proliferation curves, whatever are the considered frequency of culture medium changes and serum concentrations. Thus, the model enables an intrinsic quantification of the parameters driving in vitro EAhy926 proliferation, including proliferation, nutrient consumption and toxicity increase rates, rather independently of the experiments design. We therefore propose that such models could provide a basis for a standardized quantification of intrinsic cell proliferation kinetics.
CNRS, Laboratoire TIMC, Equipe Dynacell, Institut de l'Ingénierie et de l'Information de Santé (InS), Faculté de Médecine, 38706 La Tronche Cedex, France. philippe.tracqui@imag.fr
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P
Tracqui
J W
JW
Liu
O
O
Collin
J
J
Clement-Lacroix
E
E
Planus
D002452:Cell Count / Q000379:methods* / Q000592:standards; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology*; D045744:Cell Line, Tumor; D003470:Culture Media / Q000633:toxicity*; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D006863:Hydrogen-Ion Concentration; D008175:Lung Neoplasms; D008954:Models, Biological; D015203:Reproducibility of Results; D014471:Umbilical Veins / Q000166:cytology
D003470:Culture Media