Culture of Human Laryngeal Papilloma Cells in Vitro

Human laryngeal epithelial cells have been grown in tissue culture in a hydrated collagen gel containing Nutrient Mixture F12 (Gibco) supplemented with 15% fetal calf serum and 10μg//mL hydrocortisone. Primary cells often remain viable in culture for more than six months. They can be serially transferred two to four times before senescence. Cells derived both from normal epithelium and from laryngeal papilloma have been successfully cultured. Papilloma cells appear to contain more perinuclear granules and form fewer tight junctions than normal cells.

Establishment and transformation diminish the ability of fibroblasts to contract a native collagen gel.

Cultures of established and transformed fibroblasts were less able to contract a hydrated collagen gel than normal precrisis cells. Postcrisis fibroblasts from different rodent strains and species underwent a further reduction in contraction ability and either spontaneous or simian virus 40 (SV40) transformation. Human precrisis fibroblasts contracted much more efficiently than two SV40-transformed human lines. Fibroblasts from a patient with Glanzmann's thrombasthenia were intermediate between all other human fibroblasts assayed and the SV40-transformed human lines. The absolute efficiency of contraction was dependent on temperature and serum concentration, but no conditions were found that resulted in equal efficiencies for the three types of cells. Precrisis cells were extremely sensitive to the passage procedures when assayed for collagen contraction.