مقایسه روش‌های آزمایشگاهی IVF و Piezo-ICSI از نظر تکوین جنین در موش NMRI غیر همخون

نویسندگان

عضو هیأت علمی موسسه تحقیقات واکسن و سرم سازی رازی

چکیده

برای انجام لقاح آزمایشگاهی IVF ، از 20 سرموش ماده نژاد غیرهمخون NMRI پس از ازدیاد تخمک گذاری، 435 اووسیت در محیط 2M حاوی 4 درصد BSA جمع آوری شد که 85 درصد (370=n) از آن ها در فاز MII بودند. تعداد 332 اووسیت در قطرک های محیط T6 حاوی 4 درصد BSA  در مجاورت اسپرم های اپیدیدیمی در انکوباتور قرارگرفتند. میزان لقاح  اووسیت ها 8/79 درصد (265=n) مشاهده و ثبت گردید که 5/27 درصد (73=n) از این سلول های لقاح یافته به مرحله جنینی مورولا، 8/20 درصد (55=n) به مرحله جنینی بلاستوسیت رسیدند برای انجام پیزو –ICSI (Piezo- ICSI)، نیز از 20 سر موش ماده نژاد NMRI سوپر اووله شده، استفاده شد. تعداد  355  اووسیت در محیط M2 حاوی 4 درصد BSA جمع آوری شدند. ارزیابی ها نشان داد 83 درصد (295=n) از اووسیت ها در فاز MII بودند. با استفاده از دستگاه میکرو انجکشن پیزو، اسپرم های اپیدیدیمی به داخل سیتوپلاسم 213 اووسیت فاز MII  تزریق و سپس اووسیت ها انکوبه شدند. مشاهدات نشان داد (133=n) 4/62 درصد از اووسیت ها پس از ریز تزریقی لقاح یافتند و 14 درصد (30=n) منحط شدند. 39 درصد (52=n) از سلول های لقاح یافته به مرحله مورولا، و 8/12 درصد (17=n) به مرحله بلاستوسیت در محیط آزمایشگاه رسیدند. در این مطالعه، میزان باروری سلول های جنسی، رشد و بقای جنین های تولید شده در محیط آزمایشگاه مورد ارزیابی قرار گرفت.  نتایج نشان می دهند که با استفاده از تکنیک لقاح آزمایشگاهی (IVF) در موش آزمایشگاهی غیرهمخون- متعارف در شرایط معمول، میزان لقاح آزمایشگاهی افزایش یافته و جنین های آزمایشگاهی با کیفیت وکمیت بهتری تولید می شوند ضمن آنکه روشی آسان تر و مقرون به صرفه تر از روش میکرواینجکشن پیزو -ICSI است. بطور کلی پیشنهاد می شود در انجام تحقیقات از موش آزمایشگاهی غیر همخون NMRI که در شرایط متعارف به آسانی قابل تکثیر و نگهداری می باشند، برای دریافت تخمک و تولید جنین در فناوری باروری یاری شده (TechnologyReproductionAssisted) استفاده گردد.

عنوان مقاله [English]

Comparison of two IVF and Piezo-ICSI experimental methods from standpoint of embryo development in outbred NMRI mouse

نویسندگان [English]

  • M.H Motedayen
  • F Todehdehghan
  • S Haghighi
Members of Scientific Board of Razi Vaccine and Serum Research Institute, Karaj Iran.
چکیده [English]

In order to do in vitro fertilization (IVF), 435 oocytes were collected from 20 super ovulated NMRI female mice, in M2 media with 4% BSA. 85% (n=370) of collected oocytes were in MII phase. Epididymal sperms were added to 332 oocytes in T6 media drops with 4% BSA and incubated. Fertilization rate was 79.8% (n=265) out of which 27.5% (n=73) grew to morula and 20.8% (n=55) achieved blastocyst stages.For Piezo- intracytoplasmic sperm injection (Piezo-ICSI), 20 super ovulated female mice were sacrificed and 355 oocytes were collected in M2 media with 4% BSA. 83% (n=295) of collected oocytes were in MII phase. Epididymal sperms were piezo-injected to cytoplasm of 213 MII oocytes and incubated. 62.4% (n=133) of injected oocytes were fertilized and 14% (n=30) degenerated. 39% (n=52) of fertilized cells grew to morula and 12.8% (n=17) to blastocyst stages embryos at laboratory conditions. In this study the fertilization rate of gametes, growth and survival rate of laboratory embryos were evaluated in vitro. Results show higher ertilization rate and the laboratory embryos with desirable quality and quantity were obtained by the use of IVF technique from conventional-outbreed laboratory mouse, that is economical and easy as compare to Piezo-ICSI. However, outbred laboratory NMRI mouse that can be easily bred and kept in conventional conditions is recommended for use in oocyte recovery and embryo production in assisted reproductive technology.

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