Changes of lung cancer stem cells in patients with lung squamous carcinoma during primary culture
10.3969/j.issn.2095-4344.2015.32.016
- VernacularTitle:人肺鳞癌来源肺癌干细胞原代培养：数量及功能变化
- Author:
Zheliang LIU
;
Jiao WU
;
Linxian WANG
;
Yuejun CHEN
;
Guanyu WU
;
Gaoming XIAO
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2015;(32):5172-5176
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Studies have shown that lung cancer stem cels can be isolated from the lung cancer cel lines, But there are few reports on in vitro isolation, culture and identification of lung cancer stem cels in patients with lung squamous carcinoma.
OBJECTIVE:To establish the feasible methods of harvesting lung cancer stem cels from fresh lung cancer tissues in patients with lung squamous carcinoma, and to investigate the alterations in cel number and function during primary culture.
METHODS: Side population cels were isolated by colagenase digestion, Ficol density gradient centrifugation and Hoechst 33342 efflux properties. The isolated cels were isolated and cultured in conditioned medium. Flow cytometry method was used to detect lung cancer stem cels based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+ and CD133+/CD44+ were recorded. The single cel clones assay, flat colony formation assay and the cel sphere formation assay were used to identify the stem-like characteristics of lung cancer stem cels between the first and fourth generations.
RESULTS AND CONCLUSION:The positive rates of CD133+, CD44+ and CD133+/CD44+ cels at the fourth generation were increased significantly, and the positive rates of CD133+ and CD133+/CD44+ cels at passage 4 were significantly higher than those at the first generation. The abilities of single cel clone formation, the flat colony formation and the cel sphere formation in the fourth-generation cels were greatly enhanced compared with the first-generation cels. Experimental findings showed that stem cel-like lung cancer cels were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which stably and rapidly amplified in vitro, laying the foundation for the further study of the heterogeneity and drug resistance of lung cancer stem cels.
