The Ukrainian Biochemical Journal (Feb 2021)

Comparative characteristic of lung cancer stem-like cells generated in vitro under different culture conditions

  • O. V. Skachkova,
  • O. I. Gorbach ,
  • M. V. Inomistova,
  • L. V. Garmanchuk,
  • N. M. Khranovska

DOI
https://doi.org/10.15407/ubj93.01.088
Journal volume & issue
Vol. 93, no. 1
pp. 88 – 95

Abstract

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Cancer stem cells (CSCs) play an important role in resistance to cancer treatment and recurrence developing. The aim of this study was to obtain cell culture of MOR line non-small cell lung cancer cells enriched in CSCs and to investigate its functional and molecular-genetic properties. Tumor spheroids (TS) of MOR cell line were generated in vitro under normal adhesive (0.2% carboxymethyl cellulose, CMC) or low-adhesive (2% agarose) culture conditions. Lateral population of TS was evaluated by flow cytometry with the use of R-123 fluorescent dye, the index of R-123 exclusion was also assessed. Expression of CD44, ALDHA1, CD133, Sox2 and Nanog mRNA was determined with RT-qPCR. It was found that regardless of the culture conditions tumor spheroids form a lateral population characterized by an increased dye exclusion index. Expression levels of CD44, ALDHA1, CD133, Sox2 and Nanog mRNA in TS cells obtained under low-adhesive (2% agarose) conditions were significantly higher than in monolayer cells and cells obtained using 0.2% CMC. Thus, the proposed method of culturing in low-adhesive conditions allowed to enrich significantly tumor spheroids of MOR line in cells with CSC properties.

Keywords