미성숙난자의 형태학적 지표와 체외성숙능과의 관계

Morphologic Parameters and in vitro Maturational Competence of Human Immature Oocyte Obtained from Stimulated IVF Cycle

  • 안소정 (분당서울대학교병원 산부인과) ;
  • 지병철 (분당서울대학교병원 산부인과) ;
  • 문정희 (분당서울대학교병원 산부인과) ;
  • 이정렬 (분당서울대학교병원 산부인과) ;
  • 서창석 (분당서울대학교병원 산부인과) ;
  • 김석현 (서울대학교병원 산부인과)
  • An, So-Jung (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Jee, Byung-Chul (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Moon, Jeong-Hee (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Lee, Jung-Ryeol (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Suh, Chang-Suk (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Kim, Seok-Hyun (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University)
  • 발행 : 2008.12.30

초록

목 적: 미성숙 난자의 체외성숙에 있어 난구세포의 양상 및 미성숙 난자의 크기가 영향 인자로 작용하는지를 알아보고자 하였다. 연구방법: 체외수정을 위한 과배란유도 후 난자채취를 시행한 21명의 환자로부터 41개의 난포 단계의 난자를 얻었다. Denudation 이전의 난구세포의 모양에 따라 dispersed 난구세포와 compacted 난구세포로 나누었다. Denudation 이후 투명대를 포함하는 난자외직경 (outer oocyte diameter)과 투명대를 포함하지 않는 난자내직경 (inner oocyte diameter)을 측정하고 체외성숙 배양액에서 체외성숙을 시도하였다. 난자의 성숙은 제1극체가 나온 경우로 정하였다. 성숙이 확인된 난자는 ICSI 방법을 이용하여 수정시키고 다음날 두 개의 전핵이 뚜렷이 보이는 경우에 수정된 것으로 간주하였다. 결 과: 전체적인 체외성숙률은 56.1%, 수정률은 73.9%이었다. Dispersed 난구세포 난자 중에서 성숙된 난자 비율 (91.3%)은 비성숙 난자 비율 (55.6%)보다 유의하게 높았다 (p=0.023). 성숙 또는 비성숙 난자에서 난자외경($155.7{\mu}m$ vs. $152.4{\mu}m$, NS)과 내경 ($114.3{\mu}m$ vs. $113.4{\mu}m$, NS) 및 투명대 두께 ($41.3{\mu}m$ vs. $39.1{\mu}m$, NS)는 차이가 없었다. 체외성숙률은 dispersed 난구세포 난자에서 compacted 난구세포 난자보다 의미있게 높았으나 (67.7% vs. 20.0%, p=0.044), 수정률에 있어 두 군간의 유의미한 차이는 없었다. 난자의 외경, 내경 및 투명대 두께에 따른 체외성숙률 및 수정률의 차이가 없었다. 결 론: Dispersed 난구세포를 가진 미성숙 난자가 compacted 난구세포를 가졌던 난자에 비하여 더 높은 체외수정능을 가진다는 결과는 dispersed 난구세포가 미성숙 난자의 체외성숙 예측을 위한 하나의 지표로 사용될 수 있음을 시사한다.

Objective: This study was performed to investigate whether cumulus morphology and oocyte diameter influence on in vitro maturation (IVM) of human germinal vesicle (GV) stage oocytes obtained from stimulated in vitro fertilization (IVF) cycles. Methods: Forty-one GV stage oocytes were obtained from 21 patients who received ovarian hyperstimulation and IVF. According to cumulus morphology before denudation, GV oocytes were classified into oocytes with dispersed cumulus cells (CCs) or compacted CCs. The diameters of denuded oocytes, both including and excluding the zona pellucida, were measured. All oocytes were cultured in commercial IVM medium. Maturation was defined as extrusion of the first polar body and the matured oocytes were inseminated by ICSI method. Results: Overall maturation and fertilization rate were 56.1% and 73.9%. Matured oocytes had significantly higher proportion of oocytes with dispersed CCs compared to oocytes failed to mature (91.3% vs. 55.6%, p=0.023). There were no significant differences of oocytes outer ($155.7{\mu}m$ vs. $152.4{\mu}m$, NS), inner ($114.3{\mu}m$ vs. $113.4{\mu}m$, NS) diameters and zona thicknesses ($41.3{\mu}m$ vs. $39.1{\mu}m$, NS) between matured and not-matured oocytes. In-vitro maturation rate of oocytes with dispersed CCs was significantly higher than which of oocytes with compacted CCs (67.7% vs. 20.0%, p=0.044). Oocyte diameters (outer and inner) and thicknesses were not related with maturational competence. Conclusion: Our results suggest that in vitro maturational competence of GV stage oocytes obtained from stimulated IVF cycles is closely associated with the cumulus morphology but not oocyte diameter.

키워드

참고문헌

  1. Chian RC. In-vitro maturation of immature oocytes for infertile women with PCOS. Reprod Biomed Online 2004; 8(5): 547-52 https://doi.org/10.1016/S1472-6483(10)61101-7
  2. Jee BC, Han SH, Moon JH, Suh CS, Kim SH. Influence of well-defined protein source on in vitro maturation of human oocyte: human follicular fluid versus human serum albumin. Fertil Steril 2008; 89: 348-52 https://doi.org/10.1016/j.fertnstert.2007.02.052
  3. Lim JH, Yang SH, Chian RC. New alternative to infertility treatment for women without ovarian stimulation. Reprod Biomed Online 2007; 14(5): 547-9 https://doi.org/10.1016/S1472-6483(10)61042-5
  4. Holzer H, Scharf E, Chian RC, Demirtas E, Buckett W, Tan SL. IVM of oocytes collected from unstimulated ovaries for oocyte donation. Fertil Steril 2007; 88(1): 62-7 https://doi.org/10.1016/j.fertnstert.2006.11.087
  5. Huang JY, Tulandi T, Holzer H, Tan SL, Chian RC. Combining ovarian tissue cryobanking with retrieval of immature oocytes followed by IVM and vitrification: an additional strategy of fertility preservation. Fertil Steril 2008; 89(3): 567-72 https://doi.org/10.1016/j.fertnstert.2007.03.090
  6. Oktay K, Demirtas E, Son WY, Lostritto K, Chian RC, Tan SL. In vitro maturation of germinal vesicle oocytes recovered after premature luteinizing hormone surge: description of a novel approach to fertility preservation. Fertil Steril 2008; 89: 228.e19-22
  7. Heindryckx B, De Sutter P, Gerris J, Dhont M, Van der Elst J. Embryo development after successful somatic cell nuclear transfer to in vitro matured human germinal vesicle oocytes. Hum Reprod 2007; 22(7): 1982-90 https://doi.org/10.1093/humrep/dem106
  8. Jurema MW, Nogueira D. In vitro maturation of human oocytes for assisted reproduction. Fertil Steril 2006; 86: 1277-91 https://doi.org/10.1016/j.fertnstert.2006.02.126
  9. Wynn P, Piclon HM, Krapez JA. Pretreatment with follicle stimulating hormone promotes the number of human oocytes reaching metaphase II by in-vitro maturation. Hum Reprod 1998; 13: 3132-8 https://doi.org/10.1093/humrep/13.11.3132
  10. Sato C, Shimada M, Mori T, et al. Assessment of human oocyte developmental competence by cumulus cell morphology and circulating hormone profile. Reprod Biomed Online 2007; 14: 49-56 https://doi.org/10.1016/S1472-6483(10)60763-8
  11. Yang SH, Son WY, Yoon SH, Ko Y, Lim JH. Correlation between in vitro maturation and expression of LH receptor in cumulus cells of the oocytes collected from PCOS patients in HCG-primed IVM cycles. Hum Reprod 2005; 20(8): 2097-103 https://doi.org/10.1093/humrep/dei045
  12. Choi J, Kim D, Cha J, Lee S, Yoon S, Lim J. The relationship between oocyte size and developmental competence in unstimulated cycles. ASRM 2006 P-705
  13. Cavilla JL, Kennedy CR, Byskov AG, Hartshorne GM. Human immature oocytes grow during culture for IVM. Hum Reprod 2008; 23(1): 37-45
  14. Loutradis D, Kiapekou E, Zapanti E, Antsaklis R. Oocyte Maturation in Assisted Reproductive Techniques. Ann NY Acad Sci 2006; 1092: 235-46 https://doi.org/10.1196/annals.1365.020
  15. Trouson A, C. wood, A. Kausche. In vitro maturation and the fertilization and developmental competence of oocytes recovered from untreated polycystic ovarian patients. Fertil Steril 1994; 62: 353-62 https://doi.org/10.1016/S0015-0282(16)56891-5
  16. Arlotto T, Schwartz JL, First NL, Leibfried-Rutledge ML. Aspects of follicle and oocyte stage that affect in vitro maturation and development of bovine oocytes. Theriogenology 1996; 45: 943-56 https://doi.org/10.1016/0093-691X(96)00024-6
  17. Raghu HM, Nandi S, Reddy SM. Follicle size and oocyte diameter in relation to developmental competence of buffalo oocytes in vitro. Reprod Fertil Dev 2002; 14: 55-61 https://doi.org/10.1071/RD01060
  18. Anguita B, Vandaele L, Mateusen B, Maes D, Van Soom A. Developmental competence of bovine oocytes is not related to apoptosis incidence in oocytes, cumulus cells and blastocysts. Theriogenology 2007; 67(3): 537-49 https://doi.org/10.1016/j.theriogenology.2006.09.004
  19. Anguita B, Jimenez-Macedo AR, Izquierdo D, Moga T, Paramio MT. Effect of oocyte diameter on meiotic competence, embryo development, p34 (cdc2) expression and MPF activity in prepubertal goat oocytes. Theriogenology 2007; 67(3): 526-36 https://doi.org/10.1016/j.theriogenology.2006.09.003
  20. Hamamah S. Oocyte and embryo quality: is their morphology good criterion? J Gynecol Obstet Biol Reprod (Paris) 2005; 34: 5S38-41
  21. Serhal PF, Ranieri DM, Kinis A, Marchant S, Davies M, Khadum IM. Oocyte morphology predicts outcome of intracytoplasmic sperm injection. Hum Reprod 1997; 12: 1267-70 https://doi.org/10.1093/humrep/12.6.1267
  22. Xia P. Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality. Hum Reprod 1997; 12: 1750-5 https://doi.org/10.1093/humrep/12.8.1750
  23. Wilding M, Matteo LD, D'Andretti S, Montanaro N, Capobianco C, Dale B. An oocyte score for use in assisted reproduction. J Assist Reprod Genet DOI 10.1007/s10815-007-9143-8