Establishment and phenotypic characterization of the first human pulmonary blastoma cell line

Lung Cancer. 2011 Apr;72(1):23-31. doi: 10.1016/j.lungcan.2010.07.009. Epub 2010 Aug 17.

Abstract

Background: Sarcomatoid carcinomas are a group of poorly differentiated carcinomas containing a sarcomatoid component with the presence of epithelial-mesenchymal transition.

Materials and methods: To obtain a stabilized cell line, three mediums were used: IMDM, BEBM and IMDM/BEBM. CD44, CD29, CD90, CD133, CD326 antigens, cytokeratins and vimentin were tested by flow cytometry and immunofluorescence assay. Side population, spheres formation, tumorigenicity in vitro and in vivo and cell cycle analysis were analyzed.

Results: At day of surgery, cytometric analysis revealed that CD133, CD90 and CD326 levels were very low, CD29 and CD44 were about 80%. After 30 days of culture, CD133 levels increased up to 30%, all cells expressed CD90 and vimentin markers and CD326 was lost. The cells grew only in IMDM/BEBM medium. The cell population was heterogeneous with epithelial and mesenchymal cells. After about 15-30 days, only fibroblast-like cells were observed. LC114 cells were not able to grow as spheres and they did not show a side-population phenotype. The cell cycle analysis confirmed an aneuploid population in the tissue and normal diploid population in cell line.

Conclusions: We selected, characterized and stabilized a primary cell line of human pulmonary blastoma by the expression both of stemness and differentiation markers and confirmed the presence of the marker CD133.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged, 80 and over
  • Animals
  • Antigens, CD / metabolism
  • Cell Cycle / physiology
  • Cell Line, Tumor*
  • Female
  • Humans
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology*
  • Male
  • Mice
  • Mice, Nude
  • Mice, SCID
  • Pulmonary Blastoma / metabolism
  • Pulmonary Blastoma / pathology*
  • Spheroids, Cellular
  • Transplantation, Heterologous
  • Tumor Cells, Cultured

Substances

  • Antigens, CD