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
PY - 2013/3/1
Y1 - 2013/3/1
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 -