Culturing Hi-5 and Sua5B Insect Cell Lines

We used the BTI-Tn-5B1-4 cell line [High Five Cells (Hi-5)], which is derived from ovarian cells of the cabbage looper (Trichoplusia ni) and is the standard insect cell line commonly used in many laboratories. We also chose to use the An. gambiae Sua5B cell line as these cells are immunocompetent and hemocyte-like. The Hi-5 and Sua5B cells were cultured in Express Five SFM medium (Invitrogen) and Grace’s Supplemented Insect Medium (Gibco, Waltham, MA), respectively, at 27 °C, as described previously [22 (link)]. The Sua5B cell medium also contained 10% heat-inactivated fetal bovine serum (Gibco); the cell lines were generally passaged twice per week upon reaching 90% confluency.