Evaluation of Cell Doubling Time in C6 and Y79 Cell Lines Based on Seeding Density
Shreyas S Kuduvalli, O Ramalakshmi, S Daisy Precilla, Thirugnanasambandhar S Anitha
Keywords :
Cell doubling time, Glioblastoma multiforme, Growth curve, Retinoblastoma
Citation Information :
Kuduvalli SS, Ramalakshmi O, Precilla SD, Anitha T S. Evaluation of Cell Doubling Time in C6 and Y79 Cell Lines Based on Seeding Density. 2019; 2 (4):146-149.
Aim: The purpose of this study is to evaluate the effect of seeding density on the growth pattern and measure the growth curve-related characteristics of two different central nervous system (CNS) tumor cells, i.e., C6 glioma cells and Y79 retinoblastoma (RB) cells. Materials and methods: The cell growth curve and doubling time (DT) of C6 and Y79 cells were determined by seeding 2,000, 4,000, 8,000, 16,000, and 24,000 cells/well in a 96-well plate and were incubated for different time periods such as 24, 48, 72, 96, and 120 hours. The cells were counted in a hemocytometer using a trypan blue staining method, and the optimum seeding concentration was established. Results: In this study, we observed that both the cell lines exhibited an exponential growth pattern at seeding concentrations of 2,000 and 4,000 through the incubation time of 120 hours. Interestingly, 8,000 and 16,000 cell densities reached stationary growth phase after 72 hours of exponential growth. However, at 24,000 cell density, the cells grew exponentially for just 48 hours before entering a stationary phase till 96 hours, beyond which cell death was observed with reduced cell count. Conclusion: This study implicates that both C6 and Y79 cells grow best when seeded at 4,000 cells/cm2 displaying a perfect growth curve. Furthermore, the DT for both the cell lines was observed at 24–28 hours.
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