اثرات هورمون های تحریک کننده فولیکول نو ترکیب و HMG بر بلوغ اووسیت های نابالغ انسانی در محیط کشت و میزان جنین های 8 سلولی حاصل از تزریق درون سیتوپلاسمی اسپرم

نوع مقاله: اصیل پژوهشی

نویسندگان

1 استادیار گروه زیست شناسی دانشکده علوم پایه دانشگاه آزاد اسلامی مشهد

2 استادیار دانشگاه علوم پزشکی مشهد ، مرکز تحقیقاتی و درمانی ناباروی منتصریه

3 متخصص علوم آزمایشگاهی، مرکز تحقیقاتی و درمانی ناباروری منتصریه

4 کارشناس ارشد زیست شناسی سلولی تکوینی

چکیده

مقدمه: استفاده از روش بلوغ تخمک ها در محیط کشت (IVM)برای درمان ناباروری در زنان دارای سندرم پلی کیستیک تخمدان ( PCOS) و یا افراد مبتلا به سندرم تحریک بیش از حد تخمدان (OHSS) مفید می باشد و باعث کاهش میزان مصرف داروهای گنادوتروپین و در نتیجه کاهش هزینه های درمان می گردد. مطالعه حاضر با هدف بررسی اثرات هورمون های HMG و r-FSH بر بلوغ اووسیت های نابالغ انسانی و میزان جنین های هشت سلولی ایجاد شده ، حاصل از تزریق اسپرم به سیتوپلاسم تخمک ( ICSI) انجام شده است.
 
روش کار: این مطالعه از نوع تجربی و در سال 1386-1385 در مرکز تحقیقاتی درمانی ناباروری منتصریه و دانشگاه آزاد اسلامی مشهد انجام شده است. تعداد 156 تخمک نابالغ حاصل از تخمدان زنان کاندید لقاح درمحیط کشت (IVF) به سه گروه تقسیم شد. در گروه شاهد به محیط کشت اووسیت آلبومین سرم انسانی10% و در محیط کشت گروه های تجربی 1 و 2 به ترتیب r-FSH وHMG به مقدارIU/ml 075/0 اضافه شد. سپس تخمک های بالغ را با تزریق اسپرم به داخل سیتوپلاسم آنها بارور وتعداد رویان های هشت سلولی ایجاد شده توسط میکروسکوپ معکوس بررسی گردید. داده های کمی حاصل با استفاده از روش آماری مجذور کای و نرم افزار SPSS در سطح معنی داریP<0.05 تحلیل گردید.
 
نتایج: میزان بلوغ اووسیت های نابالغ انسانی تا مرحله متا فاز 2 و رسیدن رویان ها به مرحله هشت سلولی به طور معنی داری در گروه های HMG و r-FSH نسبت به گروه کنترل افزایش نشان دادP<0.001) (.
 
نتیجه گیری:هردو هورمونHMG و r-FSH باعث افزایش میزان بلوغ اووسیت های نابالغ انسانی در محیط کشت می شود ونیز میزان رسیدن رویان ها را به مرحله هشت سلولی افزایش می دهند. 

کلیدواژه‌ها


عنوان مقاله [English]

Effects of Human Menopausal Gonadotropin and Recombinant Follicle Stimulating Hormones on In Vitro Maturation of Human Immature Oocytes and Rate of Eight-Cell Embryos Mediated by Intracytoplasmic Sperm Injection

نویسندگان [English]

  • Javad Baharara 1
  • Nozhat Mousavi fard 2
  • Mohsen Jalali 3
  • Maryam Maghani 4
1 Assistant Professor of Biology, Faculty of Sciences, Islamiz Azad University, Mashhad, Iran
2 Assistant Professor, Montaserieh Research & Clinical Center for Infertility, Mashhad University of Medical Sciences, Mashhad, Iran
3 Ph.D., Montaserieh Research & Clinical Center for Infertility, Mashhad University of Medical Sciences, Mashhad, Iran
4 M.Sc. in Biology, Faculty of Sciences, Islamiz Azad University, Mashhad, Iran
چکیده [English]

Introduction: Treatment of infertility by using in vitro matured oocytes derived from pre-ovulatory follicles has many potential applications. It minimizes the risk of ovarian hyperstimulation syndrome and is an alternative treatment for polycystic ovary syndrome.It also reduces costs and duration of treatment and can be important for subjects who need preservation. So provides information about final stage of oocyte maturation. We aimed to study the effects of recombinant follicle stimulating hormone (r-FSH) and human menopausal gonadotropin hormone (HMG) on human immature oocyte maturation in culture media and evaluated rate of eight-cell embryos mediated by intracytoplasmic sperm injection (ICSI).
Material and Methods: This experimental study was done in 2006-2007 in Montaserieh IVF Center and Mashhad Islamic Azad University. One hudred fifty six immature oocytes produced by women undergoing IVF were divided to three groups. In control group the oocytes were supplemented by 10% human serum albumin. In r-FSH group the oocytes were supplemented by 0.075 IU/ml r-FSH. And in the HMG group the oocytes were supplemented by 0.075 IU/ml HMG. Then matured oocytes were fertilized by ICSI and the numbers of embryos up to eight cells were investigated by invert microscope.
Results: The precentage of oocytes reaching metaphase 2 and the rate of eight-cell embryos was significantly higher in the FSH and HMG groups compared with the control group (p<0.001).
Conclusion: Using either HMG or r-FSH can increase the rate of human immature oocytes maturation in vitro. HMG significantly increased the production rate of eight-cell embryos compared with r-FSH (p<0.001).
 

کلیدواژه‌ها [English]

  • IVM
  • HMG
  • FSH
  • ICSI
1. Jaroudi KA , Hollanders JMG , Elnour AM , Roca GL , Atared AM, Coskun S, et al. . Embryo
development and pregnancies from in–vitro matured and fertilized human oocytes,Hum Reprod
1999 Jul;14(7):1749-51.
2. Hwu1 YM, Lee1 RK, Chen CP, Su JT, Chen YW, Lin SP.Development of hatching blastocysts
from immature human oocytes following in-vitro maturation and fertilization using a co-culture
system .Hum Reprod 1998 Jul;13(7):1916–21.
3. Pincus G, Enzmann EV.The comparative behavior of mammalian eggs in vivo and in vitro. I The
activation of ovarian eggs. J Exp Med 1935; 62: 665-75.
4. Combelles CM, Cekleniak NA, Racowsky C, Albertini DF.Assessment of nuclear and
cytoplasmic maturation in in-vitro matured human oocytes.Hum Reprod 2002 Apr;17(4):1006-16.
5. Khashavi Z, Karimzadeh MA.Comparison of unstimulated in vitro maturation and stimulated in
vitro fertilization in women with poly cystic ovarian syndrome.J Med Science 2004;6:11-14.
6. Giudice E, Crisci C, Eshkol A and Papoian R. Composition of commercial gonadotropin
preparations extracted from human post-menopausal urine: characterization of non-gonadotropin
proteins.Hum Reprod 1994 Dec;9(12): 2291–9.
7. Accardo C, Dattena M, Pilichi S, Mara L, Chessa B and Cappai P. Effect of recombinant human
FSH and LH on in vitro maturation of sheep oocyte; embryo development and viability.Anim
Reprod Sci 2004 Mar;81(1-2):77-86.
8. Anderiesz C, Ferraretti A, Magli C, Fiorentino A, Fortini D, Gianaroli L, et al. Effect of
recombinant human gonadotrophins on human, bovine and murine oocyte meiosis, fertilization
and embryonic development in vitro. Hum Reprod 2000 May;15(5):1140-8.
9. Zhang X, Zerafa A, Wong J, Armstrong DT, Khamsi F. Human menopausal gonadotropin during
in vitro maturation of human oocytes retrieved from small follicles enhances in vitro fertilization
and cleavage rates. Fertil Steril 1993 Apr;59(4):850-3.
10. Daya S, Gunby J, Hughes EG, Collins JA, Sagle MA. Follicle-stimulating hormone versus human
menopausal gonadotropin for in vitro fertilization cycles.Fertil Steril 2001 Oct ;76(4):853-4.
11. Imthurn B, Macas E, Rosselli M, Keller PJ. Nuclear maturity and oocyte morphology after
stimulation with highly purified follicle stimulating hormone compared to human menopausal
gonadotrophin. Hum Reprod 1996 Nov;11(11):2387-91.
 
12. Agrawal R, Holmes J, Jacobs HS. Follicle-stimulating hormone or human menopausal
gonadotropin for ovarian stimulation in in vitro fertilization cycles: a meta-analysis. Fertil Steril
2000 Feb;73(2):338-43.
13. Ng EH, Lau EY, Yeung WS, Ho PC. HMG is as good as recombinant human FSH in terms of
oocyte and embryo quality: a prospective randomized trial. Hum Reprod 2001 Feb;16(2):319-25.
14. Son WY, Yoon SH, Park SJ, Yoon HJ, Lee WD, Lim JH. Ongoing twin pregnancy after
vitrification of blastocysts produced by in-vitro matured oocytes retrieved from a woman with
polycystic ovary syndrome: Case report. Hum Reprod 2002 Nov;17(11):2963-6.
15. Smith GD. In vitro maturation of oocytes. Curr Womens Health Rep 2001 Oct;1(2):143-51.
16. Krisher RL. The effect of oocyte quality on development.J Anim Sci 2004;82 E-Suppl:E14-23.
17. Balakier H, Sojecki A, Motamedi G, Librach C. Time-dependent capability of human oocytes for
activation and pronuclear formation during metaphase II arrest. Hum Reprod 2004 Apr;19(4):982-
7.
18. Trounson A, Anderiesz C, Jones G. Maturation of human oocytes in vitro and their developmental
competence. Reproduction 2001 Jan;121(1):51-75.
19. Combelles CM, Albertini DF. Assessment of oocyte quality following repeated gonadotropin
stimulation in the mouse. Biol Reprod 2003 Mar;68(3):812-21.
20. Mikkelsen AL, Lindenberg S. Benefit of FSH priming of women with PCOS to the in vitro
maturation procedure and the outcome: a randomized prospective study. Reproduction 2001
Oct;122(4):587-92.
21. Zhang X, Zerafa A, Wong J, Armstrong DT, Khamsi F. Human menopausal gonadotropin during
in vitro maturation of human oocytes retrieved from small follicles enhances in vitro fertilization
and cleavage rates. Fertil Steril 1993 Apr;59(4):850-3.
22. Mercan R, Mayer JF, Walker D, Jones S, Oehninger S, Toner JP, et al. Improved oocyte quality is
obtained with follicle stimulating hormone alone than with follicle stimulating hormone/human
menopausal gonadotrophin combination. Hum Reprod 1997 Sep;12(9):1886-9.
23. Andersen AN, Devroey P, Arce JC.Clinical outcome following stimulation with highly purified
HMG or recombinant FSH in patients undergoing IVF: a randomized assessor-blind controlled
trial. Hum Reprod 2006 Dec;21(12):3217-27.
24. Bagratee JS, Lockwood G, López Bernal A, Barlow DH, Ledger WL.Comparison of highly
purified FSH (metrodin-high purity) with pergonal for IVF superovulation. J Assist Reprod Genet
1998 Feb;15(2):65-9.
25. Zuelke KA, Brackett BG. Effects of luteinizing hormone on glucose metabolism in cumulusenclosed
bovine oocytes matured in vitro. Endocrinology 1992 Dec;131(6):2690-6.
26. Cha KY, Chian RC. Maturation in vitro of immature human oocytes for clinical use. Hum Reprod
Update 1998 Mar-Apr;4(2):103-20