title: A new Bombyx mori larval ovarian cell line highly susceptible to nucleopolyhedrovirus.
abstract: Lepidopteran cell lines constitute the backbone for studying baculoviral biology in culturo and for baculovirus vector based recombinant protein expression systems. In the present study, we report establishment of a new continuous cell line designated as DZNU-Bm-1 from larval ovaries of the silkworm, Bombyx mori. The cells were grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum (FBS) and 3% heat inactivated B. mori haemolymph at 25+/-1 degrees C. A large number of attached epithelial-like and round refractive cells migrated from the explants and multiplied in the primary cultures. Both type of cells were subcultured initially for a few passages but after 10 passages the round refractive cells dominated the population, which could be subcultured continuously using MGM-448 medium with 10% FBS. The population doubling time of cell line was about 42h at 25+/-1 degrees C. The cell populations were largely diploids and triploids, while a few tetraploids and hexaploids were also observed. DNA profiles using Inter Simple Sequence Repeat (ISSR)-PCR and Simple Sequence Repeat (SSR) loci established the differences between DZNU-Bm-1 cell line and most widely used BmN cell line and the B. mori W-chromosome specific sequences confirmed the origin of DZNU-Bm-1 cell line to be from female silkworm. When cells were infected with free nonoccluded B. mori nucleopolyhedrovirus (BmNPV), the cell line was found to be highly susceptible with 92-94% of the cells harbouring BmNPV and having an average of 20-23 OBs/infected cell. We suggest the usefulness of this cell line in BmNPV based baculoviral expression system and also for studying in culturo virus replication.
authors.affiliation: Department of Zoology, Nagpur University Campus, Nagpur 440033, India. csbrnu_ngp@sancharnet.in
authors.forename: Arun M
authors.initials: AM
authors.lastname: Khurad
authors.forename: Sriramana
authors.initials: S
authors.lastname: Kanginakudru
authors.forename: Syed O
authors.initials: SO
authors.lastname: Qureshi
authors.forename: Mohan K
authors.initials: MK
authors.lastname: Rathod
authors.forename: Manoj M
authors.initials: MM
authors.lastname: Rai
authors.forename: Javaregowda
authors.initials: J
authors.lastname: Nagaraju
mesh_terms: D000818:Animals; D012831:Bombyx / Q000166:cytology / Q000254:growth & development / Q000821:virology*; D018929:Cell Culture Techniques; D002460:Cell Line / Q000821:virology*; D016172:DNA Fingerprinting; D005260:Female; D007814:Larva / Q000166:cytology; D017924:Nucleopolyhedroviruses / Q000502:physiology*; D010053:Ovary / Q000166:cytology* / Q000821:virology; D014779:Virus Replication / Q000502:physiology*