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

Camerlingo, Rosa; Franco, Renato; Tirino, Virginia; Cantile, Monica; Rocchi, Mariano; La Rocca, Antonello; Martucci, Nicola; Botti, Gerardo; Rocco, Gaetano; Pirozzi, Giuseppe

doi:10.1016/j.lungcan.2010.07.009

« Previous Next »Lung Cancer
Volume 72, Issue 1 , Pages 23-31, April 2011

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

Rosa Camerlingo
- Department of Experimental Oncology, National Cancer Institute, Via Mariano Semmola, 80131 Naples, Italy
- These authors contributed equally to this work.
Renato Franco
- Department of Pathology, National Cancer Institute, Naples, Italy
- These authors contributed equally to this work.
Virginia Tirino
- Department of Experimental Oncology, National Cancer Institute, Via Mariano Semmola, 80131 Naples, Italy
Monica Cantile
- Department of Pathology, National Cancer Institute, Naples, Italy
Mariano Rocchi
- Department of Genetics and Microbiology, University of Bari, Bari, Italy
Antonello La Rocca
- Department of Thoracic Surgery and Oncology, National Cancer Institute, Naples, Italy
Nicola Martucci
- Department of Thoracic Surgery and Oncology, National Cancer Institute, Naples, Italy
Gerardo Botti
- Department of Pathology, National Cancer Institute, Naples, Italy
Gaetano Rocco
- Department of Thoracic Surgery and Oncology, National Cancer Institute, Naples, Italy
Giuseppe Pirozzi
- Department of Experimental Oncology, National Cancer Institute, Via Mariano Semmola, 80131 Naples, Italy
- Corresponding author. Tel.: +39 0815903534; fax: +39 0815903814.

Received 3 March 2010; received in revised form 25 June 2010; accepted 18 July 2010. published online 19 August 2010.

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.

Keywords: Pulmonary blastoma, Epithelial–mesenchymal transition, CD133 marker, CD90 marker, Cancer stem cells, Lung