تاثیر سرم جنین گاوی بر رشد سلول‌های خونی لاروهایHelicoverpa armigera، Anagasta kuehniella و Arge ochropus در محیط کشت سلولی400 Ex-cell

نوع مقاله : مقاله پژوهشی


1 گروه گیاه‌پزشکی، دانشگاه گیلان، رشت، ایران

2 گروه ژنتیک و زیست‌شناسی ملکولی، دانشگاه علوم پزشکی اصفهان، ایران


استفاده از رده­های سلولی حشرات برای تولید آفت­کش­های زیستی، پروتئین­های نوترکیب یا تحویل ژن، در حال گسترش است. لذا تولید رده­های سلولی جدید به غنی­تر کردن این فن­آوری کمک می­کند، بنابراین هدف از تحقیق حاضر، تهیه چندین کشت اولیه از سلول­های خونی لاروهای کرم غوزه پنبه Helicoverpa armigera (Hubner)، شب­پره مدیترانه­ای آرد Anagasta kuehniella (Zeller) و زنبور برگ­خوار رز Arge ochropus (Gmelin) می­باشد. سلول­های خونی در محیط کشت EX-cell 400 حاوی غلظت­های مختلف 0، 5، 10، 15 و 20 درصد سرم جنینی گاوی در دمای 28 درجه سلسیوس نگه­داری شدند. غلظت­های سرم جنینی بر اساس پژوهش­های پیشین کشت سلولی حشرات انتخاب شدند. نتایج نشان داد که زمانی که سرم جنینی گاوی از محیط کشت حذف شد، هیچ رشد سلولی مشاهده نشد. بیشینه رشد سلول­های خونی لارو کرم غوزه در محیط کشت حاوی 15 درصد سرم جنینی گاوی مشاهده شد. با افزایش سرم جنینی گاوی به 20 درصد، افزایشی در رشد سلولی مشاهده نشد. بیشینه رشد سلول­های خونی لارو شب­پره مدیترانه­ای آرد در محیط کشت با 10 درصد سرم جنینی گاوی مشاهده شد. رشد این سلول­ها در سرم جنینی گاوی 15 و 20 درصد به­طور معنی­داری کاهش یافت. با افزایش غلظت سرم جنینی گاوی، رشد سلول­های خونی A. ochropus نیز افزایش یافت. در نتیجه، پژوهش حاضر، حداقل غلظت سرم جنینی گاوی مورد نیاز برای بیشینه رشد سلول­های حشرات را در محیط کشت سلولی نشان داد.


عنوان مقاله [English]

Effect of fetal bovine serum on the hemocytes growth of Helicoverpa armigera, Anagasta kuehniella and Arge ochropus in Ex-cell 400 culture

نویسندگان [English]

  • B. Valizadeh 1
  • J. Jalali Sendi 1
  • R. Salehi 2
1 Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran,
2 Department of Genetics and Molecular Biology, Medical University of Isfahan, Isfahan, Iran
چکیده [English]

The use of insect cell lines for producing bio-pesticides, recombinant protein expression or gene delivery is increasing. Hence, the establishment of new cell lines will help to enrich this technology. Therefore, the purpose of this research is preparation of several primary cultures initiated from hemocytes of Helicoverpa armigera (Hubner),Anagasta kuehniella (Zeller) and Arge ochropus (Gmelin). The cultures were incubated in EX-cell 400 medium supplemented with 0, 5, 10, 15 and 20% concentrations of fetal bovine serum (FBS) at 28ºC. The concentrations of FBS were chosen on the basis of previous research. Results showed no growth in any of the cell lines when FBS was omitted from the medium. Maximal growth of H. armigerahemocytes was obtained with 15% FBS. There was no further increase in growth as FBS concentration was increased to 20%, Maximal growth of A. kuehniella hemocytes was obtained at 10% FBS. The growth of these cells was significantly lower at 15 and 20% FBS. As the fetal bovine serum concentration increased, A. ochropus hemocytes growth increased.  In conclusion, the current investigation demonstrated the lowest concentration of FBS required for maximal growth of insect cells in vitro.

کلیدواژه‌ها [English]

  • Arge ochropus
  • Anagasta kuehniella
  • Helicoverpa armigera
  • Fetal bovine serum
  • Insect cell culture
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