Primary culture of squamous head and neck cancer with and without 3T3 fibroblasts and effect of clinical tumor characteristics on growth in vitro.

Twenty-one tumors from patients with squamous cell carcinoma of the head and neck (SCC H/N) were cultured with and without 3T3 fibroblasts in order to determine whether, on the basis of improved tissue culture medium, 3T3 cells could be deleted without altering growth and cloning efficiency. Thirty-five additional primary SCC H/N specimens, cultured in the presence of 3T3 fibroblasts, were studied to assess the effect of tumor differentiation, site of primary tumor, and site of specimen procurement on growth. The authors conclude that 3T3 cells remain essential for optimal growth and cloning efficiency. Also, 3T3 cells improved the number of successful cultures by 33% to 100% depending on the plating density, and cloning efficiency was improved by 50% in the presence of 3T3 cells. Growth did not correlate with tumor differentiation or site of origin of the tumor specimen. Culture of specimens from the primary site resulted in growth significantly more frequently than culture of specimens obtained from metastatic neck nodes.

Culture of squamous head and neck cancer on 3T3 fibroblasts following isokinetic velocity sedimentation.

Growth in culture of squamous head and neck cancer is hampered by microbial contamination, low plating efficiency, and cellular heterogeneity within tumors. Furthermore, clumps of cells must be removed if plating efficiency is to be accurately determined. Isokinetic velocity sedimentation was applied to 44 primary tumor specimens in an effort to minimize these problems. Seven fractions were evaluated for cell number, clump number, cell viability, clonogenic growth, plating efficiency, and microbial overgrowth. Unseparated specimens were simultaneously cultured. Microbial growth was significantly associated with the highest gradient fraction. Clumps were significantly associated with the lowest gradient fraction. Colony formation was significantly associated with middle gradient isokinetic velocity sedimentation, although seven specimens grew only when fractionated, suggesting the possibility of inhibitor cells within the tumor specimen.