In vitro isolation, culture and identification of lung cancer stem cells in patients with lung squamous carcinoma

doi:10.3969/j.issn.2095-4344.2015.28.018

Abstract

Abstract:

BACKGROUND: Studies have shown that lung cancer stem cells can be isolated from lung cancer cell lines. But there are few reports about in vitro isolation, culture and identification of lung cancer stem cells in patients with lung squamous carcinoma.

OBJECTIVE: To explore the feasible methods of harvesting lung cancer stem cells from fresh lung cancer tissue in patients with lung squamous carcinoma.

METHODS: Side population cells were isolated by collagenase digestion, Ficoll density gradient centrifugation and Hoechst 33342 solution. The isolated cells were suspended in conditioned medium for isolated culture. Flow cytometry method was used to detect lung cancer stem cells based on the cell surface markers CD133 and CD44, and the positive rates of CD133⁺, CD44⁺ and CD133⁺/CD44⁺ cells were recorded.

RESULTS AND CONCLUSION: Cells adhered at 0.5 hour after incubation; typical cell colony was formed at 4 days of culture; cells showed paving stone-shape at 7 days in a total number of 10⁸. The positive rates of CD133⁺, CD44⁺ and CD133⁺/CD44⁺ cells at passage 4 were increased significantly. These findings indicate that stem

cell-like lung cancer cells were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which were stably and rapidly amplified in vitro, laying the foundation for the further study on the heterogeneity and resistance of lung cancer stem cells in the future.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Lung Neoplasms, Neoplastic Stem Cells;, Phenotype, Cell Separation, Flow Cytometry

CLC Number:

R394

Liu Zhe-liang1 Wu Jiao, Wang Lin-xian, Chen Yue-jun, Wu Guan-yu, Xiao Gao-ming. In vitro isolation, culture and identification of lung cancer stem cells in patients with lung squamous carcinoma[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(28): 4526-4530.

Figures/Tables 1

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