Culture of primary lung tumors using medium conditioned by a lung carcinoma cell line

Abstract

Medium conditioned by the established lung tumor cell line A549 was used as a supplement to culture cells from primary solid lung tumors. Of 36 cases placed into culture, primary cells were obtained in 33 (91.7%). Of 29 cases in which subcultures were attempted, 18 (62.1%) were successful. Nine cell lines have been established by this technique to date. In growth assays, conditioned medium (CM) was found to stimulate both monolayer colony formation and growth in semi-solid medium of cells cultured from primary solid tumors. CM has been found to contain factors with the properties of both transforming growth factor alpha (TGF alpha) and insulin-like growth factor-I (IGF-I). The addition of a combination of these factors as purified peptides to basal medium at levels found in CM (0.1-0.5 ng/ml) stimulated colony formation of lung tumor cells by up to fourfold. These results indicate that secretion of growth factors may be important in tumor growth in vivo, and that use of CM may be a valuable tool for obtaining cultures from primary solid tumors.

The Jun Kinase 2 Isoform Is Preferentially Required for Epidermal Growth Factor-Induced Transformation of Human A549 Lung Carcinoma Cells

“…EGF stimulates the proliferation of several kinds of human tumor cells, such as gliomas (50), mammary tumor cells (1), and keratinocytes (33). Interestingly, EGF plays a major role in autocrine growth of human non-small-cell lung cancer cells (73,85) and is effective in inducing cell proliferation in lung primary tumor cells (76). The ability of many cell lines to form colonies when suspended in semisolid medium (anchorage independence) is strongly correlated with their ability to produce tumors when inoculated into animals (36,72).…”

Expression and function of the insulin‐like growth factor I system in human non‐small‐cell lung cancer and normal lung cell lines

“…We find no any significant link between observed cell behavior and histological sub-type. Cell-growth stimulation by exogenous IGF-I has also been reported for primary solid N-SCLC tumor in culture (Siegfried, 1989).…”

Cytogenetic abnormalities in non‐small cell lung carcinoma: Similarity of findings in conventional and feeder cell layer cultures

“…Tumor resections were trimmed of nontumor and necrotic tissue and minced with scalpels. Fragments of tissue were then triturated in a 10 ml pipette and plated directly onto inactivated murine Swiss 3T3 cells in 50% Ham's F-12 medium (Gibco Laboratories) containing 1% fetal bovine serum (FBS) (HyClone Laboratories, Logan, UT) and 50% basal Eagle's medium (Gibco Laboratories) containing 1 YO FBS, which was conditioned for 48 hr by the lung carcinoma cell line A549-1 (Siegfried and Owens, 1988;Siegfried, 1989). The conditioned basal Eagle's medium was centrifuged at 700 g for 10 min and filtered under positive pressure through a 0.22 pm filter prior to addition of fresh Ham's F-12 medium for use in culturing of primary tumor tissue.…”

“…ing sufficient metaphase cells from slow-growing nonsmall cell tumors. The use of the feeder cell layers combined with tumor cell-conditioned medium for production of NSCLC cell lines has been highly successful in our laboratory (Siegfried, 1987(Siegfried, , 1989Siegfried and Owens, 1988;Siegfried et al, 1991b). The establishment of these cell lines is crucial for further elucidation of the genetic events involved in the development of NSCLC.…”