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

Abstract

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.

Establishing and characterising a new cell line from Calliphora vicina (diptera: calliphoridae) fly embryonic tissues

“…C. vicina embryo cells that have undergone a series of cell duplication events acquire a fusiform shape that may be called fibroblast-like morphology. This is similar to the morphological characteristics reported for S. magellanica insect cells ( Cruz and Bello 2012 ) in lepidopteran species such as Clostera anachoreta ( Wen et al., 2009 ), Papilio demoleus ( Ding et al., 2013 ) and in Coleoptera species such as Tribolium castaneum ( Goodman et al., 2012 ).…”

Establishment and characterization of cell clones from the Papilio cell line RIRI-PaDe-3 by a high-efficiency clonal method

“…Insect cell lines are normally obtained by cutting the whole insect bodies or tissues into small pieces via mechanical force and culturing for some time. As a result, the cell lines are derived from different tissues, which give rise to remarkable differences in cell morphology and viral susceptibility (Pan et al 2010;Wen et al 2009). It is necessary to isolate the cells from heterogeneous cell populations.…”

<b>Obtenção e caracterização de células embrionárias indiferenciadas de <i>Syssphinx molina</i> (Cramer) (Lepidoptera, Saturniidae)</b>

“…Cultura de células embrionárias de Clostera anachoreta (Notodontidae) foi obtida em meio de cultura TNM-FH suplementado com soro fetal bovino (SFB) e antibióticos (WEN et al, 2009). Sohi (1995) estabelece um protocolo para Lepidoptera utilizando meio Grace de cultura de células de insetos e usou como suplementos SFB, albumina bovina, triptose e antibióticos.…”