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Volume 26 Issue 3
June 2011

    Yan-lei Wu, Lei Jiang, Yoshifumi Hashimoto, Robert R. Granados and Guo-xun Li. Establishment, Growth kinetics, and Susceptibility to AcMNPV of Heat Tolerant Lepidopteran Cell Lines[J]. Virologica Sinica, 2011, 26(3): 198-205. doi: 10.1007/s12250-011-3177-x
    Citation: Yan-lei Wu, Lei Jiang, Yoshifumi Hashimoto, Robert R. Granados, Guo-xun Li. Establishment, Growth kinetics, and Susceptibility to AcMNPV of Heat Tolerant Lepidopteran Cell Lines .VIROLOGICA SINICA, 2011, 26(3) : 198-205.  http://dx.doi.org/10.1007/s12250-011-3177-x
    shu

    建立及其生长特性和对病毒敏感性的研究

    cstr: 32224.14.s12250-011-3177-x
    • 1.

      吴艳蕾，姜磊，Yoshifumi Hashimoto,Robert RGranados,李国勋

    • 通讯作者： 李国勋, gxli@qau.edu.cn
    • 收稿日期： 2010-12-07
      录用日期： 2011-03-30

    • 目前已从鳞翅目sf-9，sf-21和一些家蚕细胞获得多株耐热细胞系，他们在染色体核型，膜脂质组成，形态和生长特性与亲代细胞系有很大的区别。本文报道了其它昆虫种类耐热细胞系的开发，测定 了它们的生长特性和对病毒敏感性。利用通过逐渐分阶段的提高温度，经2个月的处理，成功的获得了耐热细胞系，分别命名为sf9-ht33， sf9-ht35， High5-ht33， High5-ht35， MG1-ht33， MG1-ht35。现已传代70次以上，获得稳定的细胞系。结果表明，耐热细胞系的群体倍增时间比亲代细胞的短1-4小时。细胞形状没有明显变化，而sf9-ht细胞变大，High5和MG1 ht细胞变小。当细胞系在28℃，33℃，35℃，37℃下感染苜蓿银纹夜蛾核型多角体病毒时，芽殖病毒和多角体的产量在相应的细胞系中有最适宜的温度。

    Establishment, Growth kinetics, and Susceptibility to AcMNPV of Heat Tolerant Lepidopteran Cell Lines

    • 1.

      The Center for Advanced Invertebrate Cell Engineering Collaboration with BTI USA, Qingdao Agricultural University, Qingdao 266109, China

    • 2.

      Boyce Thompson Institute for Plant Research at Cornell University, Ithaca, NY 14853, USA

    • Corresponding author: Guo-xun Li, gxli@qau.edu.cn
    • Received Date: 07 December 2010
      Accepted Date: 30 March 2011

      Fund Project: Chinese National Science Foundation Project 30771451Boyce Thompson Institute for Plant Research Project BTI-QAU 1-23-2007Chinese National Basic Research Program (973) 2009CB118900

    • Lepidopteran heat-tolerant (ht) cell lines have been obtained with sf-9, sf-21 and several Bombyx cells. They have a distinct karyotype, membrane lipid composition, morphology and growth kinetics from the parental cell lines. In this paper, we report the development of ht cell lines from other insect species and examination of their growth characteristics and virus susceptibility. Adaptation of cell lines sf-9, BTI-TN-5B1-4 (High5) and BTI-TN-MG1 (MG1) to 33℃ and 35℃ was carried out by shifting the culture temperature between 28℃ and higher temperatures by a gradual stepwise increase in temperature. The process of adaption to a higher culture temperature was accomplished over a period of 2 months. The cell lines with the temperature adaption were designated as sf9-ht33, sf9-ht35, High5-ht33, High5-ht35, MG1-ht33, MG1-ht35. These cell lines have been subcultured over 70 passages. Adaption to high temperatures was confirmed by a constant population doubling time with individual cell lines. The population doubling time of heat adapted cell lines were 1-4 h less than these of parental cell lines. Cell shapes did not show obvious change, however, the cell size of sf9-ht cells was enlarged and those of High5 and MG1 ht cells were reduced after heat adaption. When the cell lines were infected with Autographa californica nuclear polyhedrosis virus (AcMNPV) at 28℃, 33℃, 35℃ and 37℃, production of budded virus and occlusion bodies in each cell line was optimum at its own adapted temperature.
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      1. Carretero M T, Carmona M J, Diez J L. 1991. Thermotolerance and heat shock proteins in Chironomus. J Insect Physiol, 37: 239-246.
          doi: 10.1016/0022-1910(91)90074-A

      2. Carvalho M G, Freitas M S. 1988. Effect of continues heat stress on cell growth and protein synthesis in Aedes albopictus. J Cell Physiol, 137: 455-461.
          doi: 10.1002/(ISSN)1097-4652

      3. Earley E M. 1975. Chromosomal preparation from monolayer cell cultures. Methods Cell Sci, 1: 31-35.

      4. Fittinghoff C M, Riddiford L M. 1990. Heat sensitivity and protein synthesis during heat-shock in the tobacco hornworm, Manduca sexta. J Comp Physiol B, 160: 349-356.
          doi: 10.1007/BF01075665

      5. Gerbal M, Fournier P, Barry P, et al. 2000. Adaptation of an insect cell line of Spodoptera frugiperda to grow at 37℃: characterization of an endodiploid clone. In Vitro Cell Dev Biol-Anim, 36: 117-124.
          doi: 10.1290/1071-2690(2000)036<0117:AOAICL>2.0.CO;2

      6. Grace T D C. 1962. Establishment of four strains of cells from insect tissues grown in vitro. Nature, 195: 788-789.
          doi: 10.1038/195788a0

      7. Grace T D C. 1967. Insect cell culture and virus research. In Vitro Cell Dev Biol – plant, 3: 104-117.
          doi: 10.1007/BF02615925

      8. Granados R R, Li G, Blissard G W. 2007. Insect cell culture and biotechnology. Virol Sin, 22: 83-93.
          doi: 10.1007/s12250-007-0010-7

      9. Granados R R, Li G, Derksen A C G, et al. 1994. A new insect cell line from Trichoplusia ni (BTI-Tn-5B1-4) susceptible to Trichoplusia ni singly enveloped nuclear polyhedrosis virus. J Invertebr Pathol, 64: 260-266.
          doi: 10.1016/S0022-2011(94)90400-6

      10. Hayflick L. 1973. Theory of population increase by subcultivation. In: Tissue Culture Methods and Application (Kruse P E & Patterrson M K. eds. ), Pulishing place: Academic Press, New York. p222-223.

      11. Imanishi S, Cho E S, Tomita S. 1999. Novel Bombyx mori cell lines cultivable at 37℃. Appl Entomol Zool, 34: 259-266.

      12. Li G X, Hashimoto Y, Granados R R. 2003. Growth characteristics and expression of recombinant protein by new cell clones derived from Trichoplusia ni (BTI-Tn-5B1-4) High Five cells. Bioprocessing, 2: 34-37.

      13. Lindquist S, Graig E A. 1988. The heat shock proteins. Annu Rev Genet, 22: 631-677.
          doi: 10.1146/annurev.ge.22.120188.003215

      14. Lynn D E. 2001. Effects of temperature on the susceptibility of insect cells to infection by baculovirus. Methods Cell Sci, 23: 221-225.
          doi: 10.1023/A:1016394421408

      15. Meng M J, Li T L, Li C Y, et al. 2008. A suspended cell lines from Trichoplusia ni (Lepidoptera): Characteri-zation and expression of recombinant proteins. Insect Sci, 15:423-428.
          doi: 10.1111/ins.2008.15.issue-5

      16. Mitchell H K, Moller G, Petersen N S. 1979. Specific protection from phenocopy induction by heat shock. Dev Genet, 1:181-192.
          doi: 10.1002/(ISSN)1520-6408

      17. Ritossa F M. 1962. A new puffing pattern induced by heat shock and DNP in Drosophila. Experimentia, 18: 571-573.
          doi: 10.1007/BF02172188

      18. Wang P, Granados R R, Shuler M L. 1992. Studies on serum-free culture of insect cells for virus propagation and recombinant protein production. J Invertebr Pathol, 59: 46-53.
          doi: 10.1016/0022-2011(92)90110-P

      19. Wang P, Toung R, Granados R R. 1999. The establishment of new cell lines from Pseudaletia unipuncta with differerntial responses to baculovirus infection. In Vitro Cell Dev Biol Animal, 35: 333-338.

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      Establishment, Growth kinetics, and Susceptibility to AcMNPV of Heat Tolerant Lepidopteran Cell Lines

        Corresponding author: Guo-xun Li, gxli@qau.edu.cn
      • 1. The Center for Advanced Invertebrate Cell Engineering Collaboration with BTI USA, Qingdao Agricultural University, Qingdao 266109, China
      • 2. Boyce Thompson Institute for Plant Research at Cornell University, Ithaca, NY 14853, USA
      • Received Date: 07 December 2010
      • Accepted Date: 30 March 2011
      Fund Project:  Chinese National Science Foundation Project 30771451Boyce Thompson Institute for Plant Research Project BTI-QAU 1-23-2007Chinese National Basic Research Program (973) 2009CB118900

      Abstract: Lepidopteran heat-tolerant (ht) cell lines have been obtained with sf-9, sf-21 and several Bombyx cells. They have a distinct karyotype, membrane lipid composition, morphology and growth kinetics from the parental cell lines. In this paper, we report the development of ht cell lines from other insect species and examination of their growth characteristics and virus susceptibility. Adaptation of cell lines sf-9, BTI-TN-5B1-4 (High5) and BTI-TN-MG1 (MG1) to 33℃ and 35℃ was carried out by shifting the culture temperature between 28℃ and higher temperatures by a gradual stepwise increase in temperature. The process of adaption to a higher culture temperature was accomplished over a period of 2 months. The cell lines with the temperature adaption were designated as sf9-ht33, sf9-ht35, High5-ht33, High5-ht35, MG1-ht33, MG1-ht35. These cell lines have been subcultured over 70 passages. Adaption to high temperatures was confirmed by a constant population doubling time with individual cell lines. The population doubling time of heat adapted cell lines were 1-4 h less than these of parental cell lines. Cell shapes did not show obvious change, however, the cell size of sf9-ht cells was enlarged and those of High5 and MG1 ht cells were reduced after heat adaption. When the cell lines were infected with Autographa californica nuclear polyhedrosis virus (AcMNPV) at 28℃, 33℃, 35℃ and 37℃, production of budded virus and occlusion bodies in each cell line was optimum at its own adapted temperature.

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