Fluorescence-based method is more accurate than counting-based methods for plotting growth curves of adherent cells

Author(s)

Pereira, Tulio Felipe ; Levin, Gabriel ; DeOcesano-Pereira, Carlos ; Caodaglio, Amanda Schiersner ; Fujita, Andre ; Tonso, Aldo ; Sogayar, Mari Cleide

• Document type: Journal article
• Source: BMC RESEARCH NOTES; v. 13, n. 1, p. 7-pg., 2020-02-04.

Abstract

Objective Cell growth curves constitute one of the primary assays employed to analyze cell proliferation dynamics of in vitro cultured cells under specific culture conditions. From the cell growth curve, it is possible to assess the behavior of proliferating cells under different conditions, such as drug treatment and genomic editions. Traditionally, growth curves for adherent cells are obtained by seeding the cells in multiple-well plates and counting the total number of cells at different time points. Here, we compare this traditional method to the fluorescence-based method, which is based on the CFSE fluorescence decay over time. Results The fluorescence-based method is not dependent on the determination of the total number of cells, but rather is approached by assessing the fluorescence of a sample of single cells from a cell population at different time points after plating. Therefore, this method is not biased due to either cell loss during harvesting or to the presence of cellular debris and cell clumps. Moreover, the fluorescence-based method displays lower variation among different measurements of the same time point, which increases the reliability on the determination of lag, log and stationary phase transitions. (AU)

• FAPESP's process: 16/05311-2 - Regenerative medicine aiming at therapy for chronic degenerative diseases (cancer and diabetes)
• Grantee: Mari Cleide Sogayar
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/01626-9 - Characterization of ENC1 (NRP/B) isoforms and identification of human glioblastoma-penetrating peptide
• Grantee: Túlio Felipe Pereira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)