Effects of hypoxia on the kinetic and morphological characteristics of human melanoma cells grown as colonies in semi-solid agar medium.
An investigation was made of the cell populations that occur in the sequential strata of human melanoma ( MM96 ) colonies. Colonies were either grown for the full duration of culture in a 'physiological' atmosphere of 5% O2 (unperturbed colonies), or grown in this atmosphere followed by a final incubation in a hypoxic atmosphere of less than 0.1% O2. Both autoradiographic and ultrastructural studies indicated that cell changes similar to those which occur successively in monolayer cultures experiencing nutritional deficiency, exist concurrently in the sequential strata of the larger unperturbed colonies. At the margin with the necrotic core, approximately half of the cells showed morphological changes associated with death by apoptosis. The other half were undergoing necrosis. Observations on colonies incubated for the final 24 or 48 h in a hypoxic (less than 0.1% O2) atmosphere showed that many of these cells, although otherwise well preserved, developed oedema complicated by cytoskeletal rupture and extrusion of areas of damaged cytoplasm within membrane-bound vesicles. Although sudden-onset hypoxia did not appear to precipitate cell death in small colonies previously lacking a necrotic core, large colonies suffered a marked reduction in the width of their viable rims. Cell death under this circumstances was by necrosis, the same mode of death as occurs with infarction. The study indicated that apoptosis was associated with sub-acute cell death as occurs with progressive nutrient depletion and catabolite accumulation, whereas necrosis was associated with acute cell death as seen in previously compromised cells subject in addition to sudden-onset hypoxia.
D000362:Agar; D000368:Aged; D002460:Cell Line; D002470:Cell Survival; D005260:Female; D006801:Humans; D007700:Kinetics; D008545:Melanoma / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009336:Necrosis; D010100:Oxygen / Q000502:physiology*; D014410:Tumor Stem Cell Assay
D000362:Agar; D010100:Oxygen