Development of a cell line from primary cultures of rainbow trout, Oncorhynchus mykiss (Walbaum), gills

Abstract. Collagenase was used to prepare primary cell cultures from rainbow trout, Oncorhynchus mykiss (Walbaum), gills. Although difficult to subcultivate, one primary culture led to the development of a cell line, RTgill‐W1. RTgill‐W1 grew in the basal medium, L‐15, supplemented with foetal bovine serum at between 5 and 10%, but not in L‐15 alone. The cells have been passaged approximately 50 times over a 4‐year period. At confluency, the cell shape was predominantly polygonal or epithelial‐like. RTgill‐Wl cultures were demonstrated by DNA staining with H33258 and by growth on agar to be contaminated with mycoplasma, but this contaminant was eradicated by treatment with mycoplasma removal agent (MRA) and BM cyclin.