ایجاد و تعیین ویژگی‌های یک رده سلولی جدید از هموسیت‌های لارو زنبور برگخوار رز Arge ochropus (Hymenoptera; Argidae)

نوع مقاله : مقاله کامل، انگلیسی

نویسندگان

1 دانشگاه گیلان

2 گروه گیاه پزشکی دانشگاه گیلان

3 گیاه پزشکی دانشگاه گیلان

4 دانشگاه علوم پزشکی اصفهان

چکیده

یک رده­ی سلولی جدید از سلول­های خونی لارو زنبور برگ‌خوار رز، Arge ochropusایجاد شد. این رده سلولی براساس نام Arge ochropus University of Guilan، AOU-G نام­گذاری شد. این رده­ی سلولی از سلول­های خونی لارو زنبور برگ‌خوار رز، به منظور ارزیابی خصوصیات و حساسیت به اکدایزون مورد بررسی قرار گرفت. ویژگی‌های شکل‌شناسی سلول‌های آن با استفاده از میکروسکوپ‌های نوری بررسی شد. از نظر ریخت شناسی در کشت اولیه سلول­های خونی، چهار نوع سلول مشهود بود که به­طور عمده شامل پروهموسیت‌ها، پلاسماتوسیت‌ها، گرانولوسیت‌ها و انوسیتوئیدها بود. زمان مضاعف شدن جمعیت این رده سلولی در بیستمین کشت مجدد در دمای 28 درجه­ی سلسیوس روی محیط کشت Ex-cell 400 و گریس به­ترتیب 3 و 5 روز بود. مطالعات بیوشیمیایی روی کشت اولیه ناشی از سلول­های خونی در محیط کشت Ex-cell 400 و گریس، نمایانگر تغییرات قابل ملاحظه­ای در برخی از ترکیبات نظیر گلوکز، پروتئین، اوره و کلسترول در محیط کشت بود. به غیر از اوره که افزایش معنی­داری را در محیط کشت نشان داد، بقیه ترکیبات اندازه­گیری شده کاهش چشمگیری پیدا نمودند. ردة سلولی AOU-G را به عنوان یک ردة سلولی جدید معرفی می­کنیم که می­تواند در مطالعات مربوط به زیست‌شناسی، ایمونولوژی، غدد درون ریز وسم­شناسی بسیار مفید باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Establishment and characterizations of a new cell line from larval hemocytes of rose sawfly Arge ochropus (Hymenoptera; Argidae)

نویسندگان [English]

  • J. Jalali-sendi 1
  • B. Valizade 2
  • R. Khosravi 3
  • R. Salehi 4
2 Plant Protection, University of Guilan
3 Plant Protection, University of Guilan
4 University of Isfahan
چکیده [English]

We established and characterized a hymenopteran cell line from Arge ochropus for the first time. The established cell line was named AOU-G, standing for Arge ochropus University of Guilan. This cell line was derived from larval hemocytes of the rose sawfly whose characteristics and susceptibility to 20-hydroxyecdysone have been verified. We also noticed four types of hemocytes such as prohemocytes, plasmatocytes, granulocytes and oenocytes. The population doubling time of 20th passage of this new cell line in EX-cell or Grace medium at 28˚C was 3 and 5 days, respectively. Biochemical studies in rose sawfly hemocytes showed considerable changes in some component: protein, glucose, cholesterol and urea. Except urea, all components were significantly reduced. We suggest that the AOU-G cell line as a productive new cell line that could be very useful in studies concerning bio- insecticides, immunology, endocrinology and toxicology. 

کلیدواژه‌ها [English]

  • Arge ochropus
  • Baculovirus
  • Cell culture
  • Ecdysone
  • Hemocytes

مراجع

Belloncik, S., Petcharawan, O., Couillard, M., Charpentier, G., Larue, B., Guardado, H., Chareonsak, S. & Imanishi, S. (2007) Development and characterization of a continuous cell line, AFKM-On-H, from hemocytes of the European corn borer Ostrinianubilalis (Hübner) (Lepidoptera, Pyralidae). In Vitro Cellular & Developmental Biology – Animal 43, 245–254.
Bergem, M., Norberg, N. & Aamodt, R. A. (2006) Long-term maintenance of in vitro cultured honey bee (Apismellifera) embryonic cells. BMC Developmental Biology 6, 17-23. doi: 10.1186/1471-213X-6-17.
Blissard, G. W. (1996) Baculovirus- insect cell interactions. Cytotechnology 20, 73 - 93.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248.
Calo, L., Fornelli, F., Nenna, S., Tursi, A., Caiaffa, M. F. &Macchia, L. (2003) Beauvericin cytotoxicity to the invertebrate cell line SF-9. Journal of Applied Genetics 44(4), 515-520.
Carlson, J. G. (1946) Protoplasmic viscosity changes in different regions of the grasshopper neuroblast during mitosis. Biological Bulletin 90, 109-121.
Cho, T., Shuler, M. &Granados, R. R. (1989) Current developments in new media and cell culture systems for the large-scale production of insect cells. Advances in Cell Culture 7, 261–277.
Fallon, A. M. & Gerenday, A. (2010) Ecdysone and the cell cycle: Investigations in a mosquito cell line. Journal of Insect Physiology 56, 1396–1401.
Gaw, Z. Y., Liu, N. T. & Zia, T. U. (1959) Tissue culture methods for cultivation of virus grasserie. Acta Virologica 3, 55-60.
Giannoulis, P., Brooks, C. L., Gulii, V., & Dunphy, G. B. (2005) Haemocytes of larval Malacosoma disstria (Lepidoptera: Lasiocampidae) and factors affecting their adhesion to glass slides. Physiology and Entomology 30, 278–286.
Goblirsch, M. J., Spivak, M. S. & Kurtti, T. J. (2013) A Cell Line Resource Derived from Honey Bee (Apismellifera) Embryonic Tissues. PLoS ONE 8 (7), 1-13. doi: 10.1371/journal.pone.0069831
Grace, T. D. (1962) Establishment of four strains of cells from insect tissues grown in vitro. Nature 195, 788-789.
Granados, R. R. & McKenna, K. A. (1995) Insect Cell Culture Methods and Their Use in Virus Research: In: Schuler, M. L., Wood, H. A., Granados, R. R., Hammer, D. A., (Eds.), Baculovirus Expression Systems and Biopesticides. Wiley Liss. New York, pp 13 -39.
Hayflick, L. (1973) Subculturing human diploid fibroblast cultures. In: Kurse, P.F., Patterson, M.K., Jr. (Eds.), Tissue Culture: Methods and Applications. Academic Press, New York, pp 220–223.
Hunter, W. B. (2010) Medium for development of bee cell cultures (Apis mellifera: Hymenoptera: Apidae). In Vitro Cellular & Developmental Biology - Animal 46, 83 - 86. doi: 10.1007/s11626-009-9246-x.
Iwabuchi, K. (2000) A continuous cell line derived from larval fat bodies of Thysanoplusia intermixta (Lepidoptera: Noctuidae). Applied Entomology and Zoology 35, 245–249.
Jones, J. C. (1959) A phase contrast study of the blood cells in Prodenia larvae (Lepidoptera). Journal of Microbiological Science 100, 17–23.
Keddie, B. A., Erlandson, M. A. & Hilchie, G. J. (1995) Establishment and characterization of three Malacosoma disstria cell lines. Journal of Invertebrate Pathology 66(2), 136-142.
Khosravi, R., Sendi, J. J., Brayner, F. A., Alves, L. C. & Feitosa, A. P. (2016) Hemocytes of the Rose Sawfly Argeochropus (Gmelin) (Hymenoptera: Argidae). Neotropical Entomology45(1), 58-65. doi: 10.1007/s13744-015-0339-9.
Matindoost, L., JalaliSendi, J., Soleimanjahi, H., Etebari, K. & Rahbarizade, F. (2008) The effects of BmNPV on biochemical changes in primary cultures of Bombyxmori embryonic tissue. In Vitro Cellular & Developmental Biology - Animal 44, 121–127. doi: 10.1007/s11626-008-9083-3.
Matindoost, L., Sendi, J. J., Soleimanjahi, H. & Etebari, K. (2006) Differences in nutrient uptake between the fat body and embryonic primary cultures of silkworm (Bombyxmori, L.). Insect Science 13, 19–24.
Meng, X., Zheng, G., Zhao, C. D, Wan, F. H. & Li, C. Y.(2017) A cell clone strain from Mythimna separata (Lepidoptera: Noctuidae) highly susceptible to Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and M. separata NPV (MsNPV). In Vitro Cellular & Developmental Biology – Animal 53(7), 646–650. doi: 10.1007/s11626-017-0166-x.
Mitsuhashi, J. (1915) TCA Manual. 1: 151-154.
Monteiro, F., Bernal, V. & Alves, P. (2017) The role of host cell physiology in the productivity of the baculovirus-insect cell system: Fluxome analysis of Trichoplusiani and Spodoptera frugiperda cell lines. Biotechnology and Bioengineering 114(3), 674–684.
Muller, H. M., Dimopoulos, G., Blass C. & Kafatos, F. C. (1999) a hemocyte-like cell line established from the malaria vector Anopheles gambiae expresses six prophenoloxidase genes. The American Society for Biochemistry and Molecular Biology 274(17), 11727-11735.
Nath, B. S., Suresh, A., Varma, B. M. & Kumar, R. P.  (1997) Changes in protein metabolism in haemolymph and fat body of the silkworm, Bombyx mori L., in response to organophosphorus insecticides toxicity. Ecotoxicology and Environmental Safety 36, 169–173.
Richmond, W. (1973) Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Journal of Clinical Chemistry and Clinical Biochemistry 19(12), 1350-1356.
Robert, R., Li, G. & Blissard, G. W. (2007) Insect Cell Culture and Biotechnology. Virologica Sinica 22(2): 83-93.
SAS institute (1997) SAS/STAT User’s Guide for Personal Computers. SAS Institute, Cary, Nc.
Sequeira, D. P.,Correia, R., Carrondo, M. J. T., Roldão, A., Teixeira, A. & Alves,P. M. (2017) Combining stable insect cell lines with baculovirus-mediated expression for multi-HA influenza VLP production. Vaccine 39, 48-52.
Siegert, K. J. (1987) Carbohydrate metabolism in Manduca sexta during late larval development. Journal of Insect Physiology 33, 421-427.
Smagghe, G., Cynthia, L. G. & Stanley, D. (2009) Insect cell culture and applications to research and pest management. In Vitro Cellular & Developmental Biology - Animal 45, 93–105.
Söderhäll, K. & Cerenius, L. (1992) Crustacean immunity. Annual Review of Fish Diseases 2, 3-23.
Steve, H. L., McIntosh, A. H., Grasela, J. J. & Goodman, C. L. (2002) The establishment pupal cell line of a Colorado potato beetle (Coleoptera: Chrysomelidae). Applied Entomology and Zoology 37(3), 447–450.
Sudeep, A. B., Shouche, Y. S., Mourya, D. T. & Pant, U. (2002) New Helicoverpa armigera Hbn cell line from larval hemocytes for baculovirus study. Indian Journal of Experimental Biology 40, 69-73.
Zheng, G. L., Li, M. M. & Li, C. Y. (2014) Establishment and characterization of three new cell lines from the embryonic tissue of Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae). In Vitro Cellular & Developmental Biology – Animal 50, 483 488. doi: 10.1007/s11626-013-9732-z.

فایل ها

هم رسانی

ارجاع به این مقاله

آمار