Two cell lines, ABADRL-Cs-W3 (W3) and ABADRL-Cs-W8A (W8), were developed from a field population of Culicoides sonorensis Wirth & Jones. The cell lines were characterized by isozyme phenotyping and the ability to support the replication of bluetongue virus (BLU) and epizootic hemorrhagic disease virus (EHDV) (Orbivirus, Reoviridae). Comparison of isozymes found in the cell lines with those found in adult C. sonorensis colony insects confirmed that the cell lines were of C. sonorensis origin. There was, however, sufficient isozyme variation present in the cell lines to construct a unique isozyme profile for each cell line. Although both cell lines supported BLU and EHDV replication to the same level, one-step growth curves for BLU indicated that virus replication was faster and attained a peak titer earlier in the W3 cell line than in the W8 cell line. Viral proteins and RNA were detected earlier in the W3 cell line as well. The accelerated virus growth kinetics observed in the W3 cell line and the adherent nature of the cells makes it more suitable for certain Orbivirus studies.