A new protocol for long-term culture of a specific subpopulation of liver cancer stem cells enriched by cell surface markers

FEBS Open Bio. 2020 Sep;10(9):1737-1747. doi: 10.1002/2211-5463.12932. Epub 2020 Jul 31.

Abstract

Liver cancer stem cells (L-CSCs) are considered to be an important therapeutic target for hepatocellular carcinoma (HCC). This study provides a new in vitro long-term culture model for a specific subpopulation of L-CSCs enriched by cell surface markers. We combined CD13, CD133 and EpCAM to selectively enrich L-CSCs, which we then cultured in modified chemically defined medium. The enriched L-CSCs exhibited enhanced proliferation, self-renewal and long-term clonal maintenance ability as compared with non-CSCs. Compared with wild-type hepatocellular carcinoma, the expression of stemness surface markers, oncogenes, drug resistance and tumorigenicity in enriched L-CSCs was significantly increased. In summary, the subpopulation of L-CSCs still maintains cancer stem cell-related phenotypes after 14 days of culture.

Keywords: cell surface markers; chemically defined medium; hepatocellular carcinoma; in vitro culture model; liver cancer stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen / metabolism*
  • Biomarkers, Tumor / metabolism*
  • CD13 Antigens / metabolism*
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology*
  • Epithelial Cell Adhesion Molecule / metabolism*
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology*
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / pathology*
  • Tumor Cells, Cultured

Substances

  • AC133 Antigen
  • Biomarkers, Tumor
  • EPCAM protein, human
  • Epithelial Cell Adhesion Molecule
  • PROM1 protein, human
  • CD13 Antigens