Two new cell lines originated from the embryos of Clostera anachoreta (Lepidoptera: Notodontidae): characterization and susceptibility to baculoviruses.

Two cell lines designated CAF-Clan I and CAF-Clan II have been established from embryos of Clostera anachoreta (Lepidoptera: Notodontidae) in TNM-FH medium containing 10% inactivated fetal bovine serum. CAF-Clan I consists of a mixture of three cell types: spherical cells, spindle-shaped cells, and giant cells. Most of the cultured cells formed a suspension in the medium and were subcultured more than 60 passages. CAF-Clan II mainly consists of spindle-shaped and spherical cells which attached to the culture surface and have undergone more than 40 passages. The cell population doubling time at 27 degrees C of CAF-Clan I at passage 22 and CAF-Clan II at passage 24 was about 68.5 and 38.2 h, respectively. The chromosome number of both cell lines at passage 15 varied from 62 to 100 in the majority of cells, though a few cells exceeded 260 (n = 30). DNA amplification fingerprinting-polymerase chain reaction analysis confirmed that the origination of the two cell lines was C. anachoreta. The susceptibility of the cell lines to baculoviruses was tested. The results showed that CAF-Clan II was susceptible to infection of Autographa californica nucleopolyhedrovirus (AcMNPV) and Ecotropis oblique nucleopolyhedrovirus (EoNPV). Occlusion bodies (OBs) production was 129 +/- 4 OBs/cell and 124 +/- 15 OBs/cell for AcMNPV and EoNPV, respectively. CAF-Clan I was less susceptible to AcMNPV compared with CAF-Clan II, while non-permissive to EoNPV.