Establishment of hanging drop 3D cell culture model in cell culture plate and comparison of cell viability detection methods
10.7501/j.issn.1674-6376.2017.08.013
- VernacularTitle:培养板悬滴法三维细胞培养模型建立及细胞活力检测方法比较
- Author:
He LIN
1
;
qiu Wan WANG
;
yuan Jia JIAO
;
shen Hao YANG
;
xiang Li REN
;
guo Li XING
Author Information
1. 沈阳化工研究院有限公司安评中心
- Keywords:
Hanging drop cell culture;
HT29 cells;
Cell viability detection;
Acid phosphatase assay (APH);
MTT;
CCK-8
- From:Drug Evaluation Research
2017;40(8):1103-1106
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.
