Characterization of a new insect cell line that is derived from the neonate larvae of Papilio xuthus (Lepidoptera: Papilionidae) and its susceptibility to AcNPV

• Published in: Tissue & cell 2012-06, Vol.44 (3), p.137-142
• Main Authors: Zhang, Xin, Feng, Ying, Ding, Wei-Feng, Chen, Xiao-Ming, Wang, Cheng-Ye, Ma, Tao
• Format: Article
• Language: English
• Subjects: AcNPV infection; Advanced Basic Science; Animals; Base Sequence; Butterflies - cytology; Butterflies - genetics; Butterflies - growth & development; Butterflies - virology; Cell Line; Characterization; Chromosomes, Insect - genetics; COI; Electron Transport Complex IV - genetics; Genes, Insect; Larva - cytology; Larva - genetics; Larva - growth & development; Mitochondria - genetics; Molecular Sequence Data; Nucleopolyhedrovirus - growth & development; Nucleopolyhedrovirus - pathogenicity; Papilio xuthus; Primary Cell Culture - methods; Time Factors; Viral Load
• ISSN: 0040-8166; 1532-3072
• DOI: 10.1016/j.tice.2011.11.007

Abstract The cell line RIRI-PX1 was established from neonate larval tissues of Papilio xuthus by performing primary cultures in the modified Grace medium that was supplemented with 20% fetal bovine serum (FBS). The cell line primarily consisted of spindle-shaped and spherical cells which attached themselves to the flask. The population-doubling times (PDTs) at the 50th and 60th passage were 42.5 h and 42.1 h respectively. The average chromosome numbers of RIRI-PX1 cell line from passage 5 to passage 50 ranged from 103 to 199. It was confirmed that RIRI-PX1 cell line was derived from P. xuthus by comparing the mitochondrial cytochrome c oxidase subunit I gene (COI) of RIRI-PX1 cells and P. xuthus eggs. This cell line was susceptible to the Autographa californica nucleopolyhedrovirus (AcNPV) and produced high yield of polyhedral occlusion bodies (43.9 OBs/cell) after 10 days of infection by AcNPV. The virus titer of AcNPV infected RIRI-PX1 cells was 3.25 × 107 TCID50 /ml. We concluded that the RIRI-PX1 cell line is established from the neonate larvae tissues successfully and the cells of the cell line are sensitive to AcNPV.