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.
Establishment and phenotypic characterization of the first human pulmonary blastoma cell line
ROCCHI, Mariano;
2011-01-01
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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
