کاربرد STO به عنوان لایه غذادهنده در کشت سلول های بنیادی اسپرماتوگونی خروس

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه علوم دامی، دانشکدۀ علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

2 استادیار گروه علوم دامی، دانشکدۀ علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

3 دانشیار گروه علوم دامی، دانشکدۀ علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

4 استادیار گروه دام و طیور، پردیس ابوریحان، دانشگاه تهران، پاکدشت

5 استادیار گروه علوم دامی، دانشکدۀ کشاورزی و منابع طبیعی، دانشگاه اراک، اراک

چکیده

در این تحقیق، اثر لایۀ غذادهندۀ STO بر کشت و تکثیر سلول‏های بنیادی اسپرماتوگونی (SSCs) خروس نابالغ در محیط آزمایشگاهی ارزیابی شد. سلول‏های بیضه، از تعداد 30 قطعه خروس نابالغ نژاد ردآیلندرد (8-4 هفته) جدا و در ظرف‏های کشت چهارخانه‏ای به‏طور جداگانه و در حضور فاکتورهای رشد bFGF و LIF در دو تیمار، سه تکرار، و پنج مشاهده در هر تکرار کشت داده شدند. کلنی‏های SSCs در روز پنجم ظاهر و در روزهای هفتم و دهم ارزیابی شدند. تعداد کلنی‏های ایجادشده، مساحت کلنی، و تعداد سلول در هر کلنی روی لایۀ غذادهنده در مقایسه با کلنی‏های کشت‏شده بدون لایۀ غذادهنده بیشتر بود (05/0≥P). نتایج ارزیابی در روز 10 کشت نشان‏دهندۀ کاهش تعداد کلنی، مساحت کلنی، و تعداد سلول/کلنی در هر دو تیمار به نسبت روز هفت کشت است. ژن C-KIT در سلول‏های تشکیل‏دهندۀ کلنی‏ها بیان نشد که می‏تواند دال بر عدم تمایز و داشتن توان خود‏نوزایی سلول‏های موجود در کلنی‏ها باشد. براساس نتایج تحقیق حاضر، استفاده از لایۀ غذادهندۀ STO در کشت و تکثیر SSCs خروس نابالغ مناسب است و توان این سلول‏ها را در تکثیر و خودنوزایی در مقایسه با کشت در تیمار بدون استفاده از لایۀ غذادهنده به‏طور بهتری حفظ می‏کند.

کلیدواژه‌ها


عنوان مقاله [English]

Applying STO as feeder layer for Rooster Spermatogonial Stem cell culture

نویسندگان [English]

  • Rasoul Karimi 1
  • Malek Shakeri 2
  • Mahdi Zhandi 2
  • Hossien Moravej 3
  • Haniyeh Banikamal 1
  • Abdollah Mohammadi-Sangcheshmeh 4
  • Mahdi Khodaei-Motlagh 5
1 M.Sc. Student, Department of Animal Science, Faculty of Agricultural Science & Engineering, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
2 Assistant Professors, Department of Animal Science, Faculty of Agricultural Science & Engineering, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3 Associate Professor, Department of Animal Science, Faculty of Agricultural Science & Engineering, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
4 Assistant Professor, Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Pakdasht, Tehran, Iran
5 Assistant Professor, Department of Animal Science, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran
چکیده [English]

The objective of this study was to evaluate the effect of the STO feeder layer on prepubertal Rhode Island Red rooster SSCs culture and proliferation in vitro. Testis cells from 30 prepubertal Rhode Island Red chicken (4-8 weeks of age), were individually separated and cultivated in the presence of  bFGF and LIF growth factors on four well plates with two treatments and three replicats and five observations per each. SSCs colonies appeared on the 5th day of culture. The number of SSCs colonies, cells/colony and colony area was measured on days 7 and 10 for both treatments. The result of the colony assay on the 7th day revealed significantly higher colony numbers as well as higher cell number/colony and colony area on the STO surface compared to colonies grown on surfaces without a feeder layer (P≤0.05). In contrast, the results of the colony assay on day 10 had declined for both treatments, as compared to day 7. Also, the C-KIT gene was not expressed which is an indication that colonies might be composed of SSCs. In conclusion, these results indicate that the use of the STO feeder layer influences the SSCs proliferation and maintenance of the prepubertal roosters in short-term culture.  

کلیدواژه‌ها [English]

  • avian spermatogonial stem cells
  • cell colony
  • cell feeder layer
  • cell growth factor
  • chicken short term culture
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