بررسی تاثیر شرایط هایپوکسی بر تکوین رویان های دوسلولی موش نژاد NMRI

نوع مقاله: مقاله پژوهشی

نویسندگان

1 پژوهشگاه ملی مهندسی ژنتیک و زیست‌فناوری (NIGEB)، تهران، ایران

2 پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری (NIGEB)، تهران، ایران

چکیده

تکوین رویان در شرایط آزمایشگاهی، وابسته به پارامتر­های متعددی است که از میان آن­ها می­توان به میزان اکسیژن محیط کشت اشاره نمود. کشت رویان در شرایط محیطی با میزان بالای اکسیژن می­تواند منجر به تولید گونه­های فعال اکسیژن شود و در نتیجه آسیب­های احتمالی را برای رویان به همراه داشته باشد. بررسی­های پیشین نشان داده است که مرحله­ی پره­نوکلئوس مرحله­ای حیاتی در نمو رویان می­باشد که نسبت به میزان اکسیژن محیط حساس است و با شروع کشت از مراحل بعدی ممکن است این اثرات مخرب اکسیژن اتمسفری ناپدید ­شود. جهت بررسی دقیق­تر این موضوع، مطالعه حاضر کشت رویان­ها از مرحله­ی دو سلولی تا مرحله­ی بلاستوسیست را مد نظر قرار داده است و کیفیت رویان­ها را در شرایط هایپوکسی (غلظت اکسیژن 5 درصد و اتمسفری (غلظت اکسیژن 20 درصد) با یکدیگر مقایسه می­کند. رویان­های دوسلولی پس از جمع­آوری، به صورت تصادفی در دو گروه هایپوکسی و اتمسفری  تقسیم و تا مرحله­ی بلاستوسیست کشت داده شدند. تاثیر غلظت متفاوت اکسیژن بر کیفیت رویان­ها با بررسی نرخ تشکیل بلاستوسیست، میزان خروج رویان از زونا، تعداد کل سلول­ها، نسبت تعداد سلول­های توده­ی داخلی بلاستوسیست به تعداد کل سلول­ها و نرخ لانه­گزینی پس از انتقال رویان ارزیابی گردید. نتایج حاکی از افزایش معنی­دار (05/0p<) در نرخ تشکیل بلاستوسیست (81/78 به 05/73)، خروج از زونا (07/56 به 50/49) و نسبت توده­ی سلولی داخلی به کل سلول­ها در رویان­های گروه هایپوکسی در مقایسه با گروه کنترل (54/0 به 28/0) بود. همچنین نرخ  لانه­گزینی پس از انتقال رویان هایپوکسی (20/59) نسبت به گروه کنترل بهبود (05/0p<) یافت (50/29). برخلاف گزارشات پیشین اثرات مخرب اکسیژن موجب سرکوب رشد رویان در تمامی مراحل نمو می­شود و براین نکته تاکید می­کند که شرایط هایپوکسی از نمو رویان بهتر حمایت می­کند.

کلیدواژه‌ها


عنوان مقاله [English]

The Effect of Hypoxia Condition on NMRI Mouse Two-cell Embryo Development

نویسندگان [English]

  • D. Bagheri 1
  • M. Dashtizad 2
  • M. Daliri 1
  • E. Hashemi 1
  • A. Rahim Tayefeh 1
1 Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB. Tehran, Iran
2 Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB. Tehran, Iran
چکیده [English]

Many factors can influence the quality of embryos produced in vitro. Oxygen concentration is one of these environment parameters, which its high level can induce the production of reactive oxygen species (ROS) causing detrimental effects on embryo. Pronuclear stage is the critical time of embryo development, affected by oxygen toxicity more than other stages, so that destructive impacts of oxygen may disappear when in vitro culture initiates from next stages. The present study was conducted to compare the effects of two-oxygen concentration, atmospheric (20% O2) and hypoxic (5% O2) on development of 2-cell embryos to the blastocyst stage. Two-cell embryos were divided randomly into 2 groups including hypoxic and atmospheric groups and were cultured to the blastocyst stage under mentioned conditions. Finally, the blastocysts quality was evaluated in terms of blastocyst formation, hatching rate, cell number and implantation rate after embryo transfer. The results showed a significant increase in the blastocyst formation, hatching and implantation rates in the hypoxic group compared to atmospheric one (P<0.05). Furthermore, total cell number and inner cell mass (ICM)/total cell number were improved in hypoxic group (P<0.05). Our results not only highlighted the fact that hypoxia condition can support embryo development better than atmospheric one, but also showed that contrary to the last reports, destructive effect of oxygen could affect the quality of mouse embryos during all developmental stages.

کلیدواژه‌ها [English]

  • Blastocyst cell number
  • Embryo culture
  • Implantation
  • Hypoxia
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