Method for retermine cell-cycle dynamics based on EdU/BrdU dual labeling
10.19405/j.cnki.issn1000-1492.2024.12.015
- VernacularTitle:基于EdU/BrdU双标记法检测细胞周期动力学的方法
- Author:
Yue YUAN
1
;
Lili AI
;
Peng ZHANG
Author Information
1. 贵州医科大学基础医学院生物学教研室,贵阳 550004;贵州医科大学组织工程与干细胞实验中心,功能核酸生物药研究重点实验室,贵阳 550004
- Publication Type:Journal Article
- Keywords:
EdU;
BrdU;
cell-cycle;
immunofluorescence;
hESC
- From:
Acta Universitatis Medicinalis Anhui
2024;59(12):2170-2175,2182
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a method for detecting the duration of the synthesis-phase(Ts)and the total cell-cycle(Tc)based on the dual labeling of 5-ethynyl-2'deoxyuridine(EdU)and 5-Bromodeoxyuridinc(BrdU).Methods HEK293T cells were double labeled with EdU and BrdU and then subjected to immunofluorescence stai-ning.The fluorescence signals were collected using a High-Content Imaging System.The numbers of EdU-positive and BrdU-positive cells in the entire cell population were counted.Based on the cell cycle calculation formula in a previously published article,it was improved to accurately calculate the duration of Ts and Tc of cells.The cell cy-cle time of human embryonic stem cells was calculated using the established method.Results Based on the kinetic parameters of the EdU/BrdU dual labeling,the Ts of human embryonic stem cells was 10.9 h,and the Tc was 15.3 h.Conclusion The EdU/BrdU dual labeling method can be used to detect the duration of the cell cycle,providing an accurate and efficient method for cell cycle analysis.
