绵羊和人卵泡液对绵羊卵母细胞体外成熟的影响。

Effect of sheep and human follicular fluid on the maturation of sheep oocytes in vitro.

作者信息

Sun F J, Holm P, Irvine B, Seamark R F

机构信息

Department of Obstetrics and Gynaecology, The University of Adelaide, South Australia, Australia.

出版信息

Theriogenology. 1994;41(4):981-8. doi: 10.1016/0093-691x(94)90513-i.

DOI:10.1016/0093-691x(94)90513-i

PMID:16727451

Abstract

This study examines the effect of sheep and human follicular fluid on the in vitro maturation (IVM) of sheep follicular oocytes. Oocyte cumulus complexes recovered post mortem were matured for 24 to 26 h at 38.6 degrees C, 5% CO(2) in air, in TCM-199 bicarbonate medium supplemented with 20% fetal calf serum (FCS) and, where stated, with maturation hormones, including FSH (5.0 microg/ml), LH (5.0 microg/ml) and estradiol (1 microg/ml), or with sheep follicular fluid recovered from large (>5 mm) or small (2 to 5 mm) ovarian follicles post mortem, or with human periovular follicular fluid obtained during routine IVF procedures. The matured oocytes were then denuded, and their maturation stage and developmental capacity were assessed by in vitro fertilization (IVF) and culture (IVC). It was found that inclusion of sheep or human follicular fluid or hormone supplements in the IVM media more than doubled the number of oocytes completing maturation (FCS alone 33%, compared with 76.2% for maturation hormones, 84.2% for fluid from large and 69.6% for fluid from small sheep follicles and 82.6% for human follicular fluid), and significantly increased fertilization rates (FCS alone 51.6%, compared with 71.9% for maturation hormones, 78.4% for fluid from the large and 75.7% for fluid from small sheep follicles and 73.1% for human follicular fluid) without discernible adverse effects on the development of the cleaving embryos to the morula or blastocyst stage in culture. Omission of FCS and supplements from the IVM medium resulted in a marked reduction (56%) in the number of oocytes maturing. This reduction could be offset to a large part, but not completely, by inclusion of human follicular fluid or human follicular fluid plus LH (5 microg/ml) in the medium. The results of this study show that addition of sheep or human follicular fluid to maturation medium can enhance rather than inhibit the maturation and fertilizability of sheep follicular oocytes in vitro.

摘要

本研究探讨绵羊和人卵泡液对绵羊卵泡卵母细胞体外成熟（IVM）的影响。死后采集的卵母细胞 - 卵丘复合体在38.6℃、含5%二氧化碳的空气中，于添加20%胎牛血清（FCS）的TCM - 199碳酸氢盐培养基中培养24至26小时，并且在有说明的情况下，添加成熟激素，包括促卵泡素（FSH，5.0微克/毫升）、促黄体素（LH，5.0微克/毫升）和雌二醇（1微克/毫升），或者添加死后从大（>5毫米）或小（2至5毫米）卵巢卵泡中采集的绵羊卵泡液，或者添加常规体外受精（IVF）程序中获得的人围卵卵泡液。然后将成熟的卵母细胞去除卵丘，通过体外受精（IVF）和培养（IVC）评估其成熟阶段和发育能力。结果发现，在IVM培养基中添加绵羊或人卵泡液或激素补充剂后，完成成熟的卵母细胞数量增加了一倍多（仅添加FCS时为33%，而添加成熟激素时为76.2%，添加大卵泡绵羊卵泡液时为84.2%，添加小卵泡绵羊卵泡液时为69.6%，添加人卵泡液时为82.6%），并且显著提高了受精率（仅添加FCS时为51.6%，添加成熟激素时为71.9%，添加大卵泡绵羊卵泡液时为78.4%，添加小卵泡绵羊卵泡液时为75.7%，添加人卵泡液时为73.1%），同时对培养中分裂胚胎发育至桑椹胚或囊胚阶段没有明显的不良影响。从IVM培养基中省略FCS和补充剂会导致成熟卵母细胞数量显著减少（56%）。通过在培养基中添加人卵泡液或人卵泡液加LH（5微克/毫升），这种减少在很大程度上但并非完全得到抵消。本研究结果表明，向成熟培养基中添加绵羊或人卵泡液可增强而非抑制绵羊卵泡卵母细胞的体外成熟和受精能力。