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. 2014 Feb 12:14:24.
doi: 10.1093/jis/14.1.24.

Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression

Gui-Ling Zheng  1 , Hong-Xu Zhou  2 , Chang-You Li  3
Affiliations

Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression

Gui-Ling Zheng et al. J Insect Sci. .

Abstract

Serumfree cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serumfree Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 ×10(6) cells/mL in serumfree culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 10(6) cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 ×10(7) TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of β-galactosidase at six days postinfection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×10(4) IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serumfree culture.

Keywords: insect cell lines; population doubling time; virus production.

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Figures

Figure 1.
Figure 1.
QB-Tn9-4s cells grown in serum-containing and serumfree media. A, QB-Tn9-4scells in TNM-FH medium. B, The early third generation QB-Tn9-4s cells in serumfree Sf-900 III medium. C, The late third generation QB-Tn9-4s cells in serumfree Sf-900 III medium. D, The fifteenth generation QB-Tn9-4s cells in serumfree Sf-900 III medium . Scale bar: 40 μm . High quality figures are available online.
Figure 2.
Figure 2.
Viability of cells cultured in TNM-FH medium and serumfree Sf-900 III media. Data in the figure are expressed as mean ± SD. High quality figures are available online.
Figure 3.
Figure 3.
Growth curves of cells cultured in TNM-FH medium and serumfree Sf-900 III media. Data in the figure are expressed as mean ± SD. High quality figures are available online.
Figure 4.
Figure 4.
Beta-galactosidase expression level of cells cultured in different media at two to eight days postinfection. High quality figures are available online.
Figure 5.
Figure 5.
Secreted alkaline phosphatase expression level in cells cultured in different media at three to nine days postinfection. High quality figures are available online.
See this image and copyright information in PMC

References

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