In Vitro Maintenance of Amoebocytes from the American Oyster (Crassostrea virginica)

Abstract

Explant and monolayer cell cultures were initiated from oyster heart and embryonic tissue and maintained for periods of a few days to 6 mo depending on the type of tissue and the culture medium. A pH of 7.0–7.3 and a temperature of 20 °C were optimum. Vertebrate cell culture media prepared in a marine saline and supplemented with fetal bovine serum and protein digests provided a suitable basal medium. Supplementation of the basal medium with oyster hemolymph or extracts of oyster tissue markedly prolonged cell maintenance. Explant cultures of heart tissue with the subsequent outward migration of individual cells were most easily initiated and maintained for periods up to 6 mo. Although several cell types were observed, actively motile, granular amoebocytes predominated. No mitotic cells were observed even following exposure to a variety of mitogens. Cultures initiated from disaggregated larvae did yield actively dividing cells. Key words: oyster, cell culture, amoebocytes