DNA Content Comparison Analysis between Living Cells and Fixed Cells in Fresh Tumor Samples
10.3321/j.issn:1000-467X.2001.05.013
- VernacularTitle:新鲜肿瘤标本活细胞与固定细胞的 DNA含量分析比较
- Author:
De-Ding TAO
1
;
Yan LENG
;
Ji-Chao QIN
;
Yuan YU
;
Jian-Ping GONG
Author Information
1. Tongji Hospital Tongji Medical College Huazhong University of Science and Technology
- From:Chinese Journal of Cancer
2001;20(5):502-504
- CountryChina
- Language:Chinese
-
Abstract:
Objective： This study was designed to establish a method of analyzing DNA content in fresh tumor using fresh cells. Methods： Clinical blood tumor samples were separated to obtain mononuclear cells by Ficoll-Hypaque, solid tumor tissues were made to be single cells by using mechanical method. The purified mononuclear cells and single cells suspensions were divided into two portions. One portion was fixed by ethanol and placed overnight, then measured DNA content by conventional flow cytometric method. The other portion was stained directly by PI （ propidium iodide) dissolved in PIPES (piperazine-Ν ,Ν '-bis[2-ethanesulfonic acid] disodium salt) buffer, then measured DNA content by flow cytometry, the DNA content results were compared with those of fixed cells. Results： There was no obvious difference in the CV（ coefficient of variation) of the peak of G0/G1 cells between fresh and fixed tumor cells from the same type tumor (t-test, P >0.05). Repetition test showed that the results of DNA content analysis using fresh cells were stable. Conclusion： Fresh tumor cells stained by PI dissolved in PIPES buffer could be used for the analysis of clinical tumor propagation and DNA ploidy.
