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

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.