Generation and characterization of non-small-cell lung cancer cell lines and clones for use in the study of immunotherapy

The in vitro study of cancer has been made easier by the use of stable tumor cell (TC) lines derived from patients to study antigen expression, immunogenicity, and response to both experimental and conventional therapeutic agents. However, the routine generation of these cell lines in some tumor histologies such as non-small-cell lung cancer (NSCLC) is difficult. In many cases, colonies of TCs do not survive, most likely due to a lack of critical growth factors in cell culture medium. Other times, TC colonies are overgrown by fibroblasts, which appear to have less stringent growth requirements. In some cases, cultures are overgrown by bacteria or mold contained in the biopsy arriving from the surgical or pathology suite. This study presents the characteristics of three new NSCLC cell lines and associated autologous clones generated from both adenocarcinoma and squamous cell carcinoma tissue. Different culture media and variable techniques were used to generate these stable TC lines. Limiting dilution analysis resulted in numerous clones, some of which displayed heterogeneity in terms of growth, antigen expression, and the ability to release cytokines. The successes and failures associated with generating TC lines are discussed in this article. Both parental cultures and related clones serve as critical reagents for the continued study of the cellular immune response to NSCLC.