Chick embryo limb bud cell culture for screening environmental contaminants

摘要

Few non-mammalian systems have been used as models for assessing the developmental toxicity of environmental contaminants although the U. S. Environmental Protection Agency recognizes mammalian in vitro systems as appropriate developmental toxicity screens. The chick embryo micromass cell culture system was tested for its predictability to screen developmental toxicants to the skeletal system. Four toxicants with known toxicities in the rodent limb bud cell culture system (arsenate, aspirin, caffeine and methylmercury) were chosen for testing in the chick culture system. Cartilage-specific products, proteoglycans, were used to determine differentiation of the cells in culture by staining with Alcian Green then measured using a spectrophotometric method. Proliferation was determined by staining with Crystal Violet. Dose and temporal response experiments were conducted to determine the most sensitive dose and time of exposure of cells in culture. Overall, the micromass cultures of the chick limb buds responded in a manner similar to those seen in the rodent culture.