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Lung cancer (LC) with its different subtypes is generally known as a therapy resistant cancer with the highest morbidity rate worldwide. Therapy resistance of a tumor is thought to be related to cancer stem cells (CSCs) within the tumors. There have been indications that the lung cancer is propagated and maintained by a small population of CSCs. To study this question we established a panel of 15 primary lung cancer cell lines (PLCCLs) from 20 fresh primary tumors using a robust serum-free culture system. We subsequently focused on identification of lung CSCs by studying these cell lines derived from 4 representative lung cancer subtypes such as small cell lung cancer (SCLC), large cell carcinoma (LCC), squamous cell carcinoma (SCC) and adenocarcinoma (AC). We identified a small population of cells strongly positive for CD44 (CD44 high ) and a main population which was either weakly positive or negative for CD44 (CD44 low/− ). Co-expression of CD90 further narrowed down the putative stem cell population in PLCCLs from SCLC and LCC as spheroid-forming cells were mainly found within the CD44 high CD90 +  sub-population. Moreover, these CD44 high CD90 +  cells revealed mesenchymal morphology, increased expression of mesenchymal markers  N-Cadherin  and  Vimentin , increased mRNA levels of the embryonic stem cell related genes  Nanog  and  Oct4  and increased resistance to irradiation compared to other sub-populations studied, suggesting the CD44 high CD90 +  population a good candidate for the lung CSCs. Both CD44 high CD90 +  and CD44 high CD90 −  cells in the PLCCL derived from SCC formed spheroids, whereas the CD44 low/−  cells were lacking this potential. These results indicate that CD44 high CD90 +  sub-population may represent CSCs in SCLC and LCC, whereas in SCC lung cancer subtype, CSC potentials were found within the CD44 high  sub-population.

Identification and Characterization of Cells with Cancer Stem Cell Properties in Human Primary Lung Cancer Cell Lines