TY - JOUR
T1 - Development and characterization of a new Bombyx mori cell line for protein expression
AU - Khurad, Arun M.
AU - Bahekar, Ravindra S.
AU - Zhang, Min Juan
AU - Tiple, Ashish D.
AU - Lee, Jae Man
AU - Zhang, Chuan Xi
AU - Kusakabe, Takahiro
N1 - Funding Information:
We are thankful to M. M. Rai and M. K. Rathod for providing healthy larvae to set up the cultures, and BmNPV infected larvae to obtain the inoculum during this study. The study was partially supported by the University Grants Commission, New Delhi to AMK (Project No. F. 31‐222/2005(SR) ).
PY - 2013/3
Y1 - 2013/3
N2 - A Bombyx mori continuous cell line, designated DZNU-Bm-17, was established from larval ovaries. The cells were initially grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum and 3% heat inactivated B. mori hemolymph at 25 ± 1 °C and later adapted gradually to TNM-FH medium. Partially adhered refractive cells were the predominant cell type in the culture. The cells took about 1055. days to complete 100 passages in TNM-FH medium. The population doubling time of the cell line was about 30-34. h at 25 ± 1 °C. The cell population was largely diploid, but a few triploids and tetraploids were also observed. DNA profiles using simple sequence repeat loci established the differences between the DZNU-Bm-1, Bm-5, DZNU-Bm-12, DZNU-Bm-17, and BmN cell lines. The cell line was susceptible to budded virus of B. mori nucleopolyhedrovirus (BmNPV), and 85-92% of the cells harbored BmNPV with an average of 15 occlusion bodies/infected cell. The cells expressed the luciferase and green fluorescent proteins using the BmNPV bacmid vector. We suggest the usefulness of the DZNU-Bm-17 cell line for BmNPV-based baculoviral expression studies.
AB - A Bombyx mori continuous cell line, designated DZNU-Bm-17, was established from larval ovaries. The cells were initially grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum and 3% heat inactivated B. mori hemolymph at 25 ± 1 °C and later adapted gradually to TNM-FH medium. Partially adhered refractive cells were the predominant cell type in the culture. The cells took about 1055. days to complete 100 passages in TNM-FH medium. The population doubling time of the cell line was about 30-34. h at 25 ± 1 °C. The cell population was largely diploid, but a few triploids and tetraploids were also observed. DNA profiles using simple sequence repeat loci established the differences between the DZNU-Bm-1, Bm-5, DZNU-Bm-12, DZNU-Bm-17, and BmN cell lines. The cell line was susceptible to budded virus of B. mori nucleopolyhedrovirus (BmNPV), and 85-92% of the cells harbored BmNPV with an average of 15 occlusion bodies/infected cell. The cells expressed the luciferase and green fluorescent proteins using the BmNPV bacmid vector. We suggest the usefulness of the DZNU-Bm-17 cell line for BmNPV-based baculoviral expression studies.
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U2 - 10.1016/j.aspen.2012.09.004
DO - 10.1016/j.aspen.2012.09.004
M3 - Article
AN - SCOPUS:84867432457
VL - 16
SP - 17
EP - 22
JO - Journal of Asia-Pacific Entomology
JF - Journal of Asia-Pacific Entomology
SN - 1226-8615
IS - 1
ER -