Cultured mussel foot cells expressing byssal protein genes

Abstract

Primary and secondary culture of isolated foot cells of the mussel Mytilus galloprovincialis was carried out with modified Leibovitz's L-15 medium containing 20% fetal calf serum, 20 mg/l kanamycin and 20 mg/l streptomycin, which was adjusted to pH 7.5 and to the osmolarity of 950 mOsM/kg. Foot cells, dissociated by a collagenase treatment, successfully attached, spread and formed a monolayer in 4 to 7 days on a dish coated with collagen type I. This culture method gave reproducible results in repeated experiments. When cells were detached with trypsin from the first culture and subcultured in a flesh medium, they adhered and flattened onto the surface of cultural dishes, and propagated into confluent monolayers within 7 days. The proliferation of cells in the primary and secondary cultures was confirmed by the measurement of DNA synthesis. By RNA blot analysis using probes for the genes encoding three byssal proteins Mgfp-1, -2 and -3, it was shown that all three genes were transcribed in the primary and secondary culture although the Mgfp-1 transcription was weak. These findings suggest that the primary and secondary culture system of mussel foot cells can be used for the in vitro study of expression of at least two byssal protein genes. J. Exp. Zool. 283:131–136, 1999. © 1999 Wiley-Liss, Inc.