Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, University of Texas Health Science Center, San Antonio, Texas 78229, USA.
Fertil Steril. 2011 Mar 15;95(4):1217-23. doi: 10.1016/j.fertnstert.2010.06.095. Epub 2010 Aug 11.
To evaluate the effects of ovarian stimulation and intracytoplasmic sperm injection (ICSI)-induced fertilization and efficacy of various culture systems on in vitro development of baboon embryos.
In vitro study, animal model.
Research laboratory.
ANIMAL(S): Baboons in laboratory animal research facility.
INTERVENTION(S): Baboons received FSH (75 IU daily) for 7 to 8 days and FSH/LH (75/75 IU daily) for 3 days, followed by hCG (2,000 IU). Oocytes were retrieved laparoscopically 36 hours after hCG. Intracytoplasmic sperm injection was performed on metaphase II (MII) oocytes. Fertilized embryos were placed into different culture conditions and feeder cell coculture. Embryo development was observed through the most advanced stages, including blastocyst formation.
MAIN OUTCOME MEASURE(S): Oocytes retrieved, fertilization rates, multicell embryo rates, and blastocyst rates.
RESULT(S): Baboon oocytes (n = 1,924, from 49 cycles) were retrieved. Significant heterogeneity was seen in ovarian response to exogenous gonadotropins and subsequent oocyte maturation. The percentage of MII oocytes showed no significant difference among individual female baboons and stimulation cycles. Nearly two thirds of MII oocytes were successfully fertilized with ICSI. Blastocyst rates varied significantly among embryos in different treatments. Coculture with feeder cells in P1/Blast, Quinn's Advantage, and Sydney IVF media generated better blastocyst rates.
CONCLUSION(S): We tested multiple media and feeder cell combinations to optimize culture conditions in baboon embryo culture and obtained a high blastocyst rate similar to those reported for rhesus monkey embryos cultured in vitro, but still lower than with assisted reproductive technologies in women.
评估卵巢刺激和胞浆内单精子注射（ICSI）诱导受精以及各种培养系统对食蟹猴胚胎体外发育的影响。
体外研究，动物模型。
研究实验室。
实验室动物研究设施中的食蟹猴。
食蟹猴接受 FSH（每天 75IU）治疗 7-8 天，然后接受 FSH/LH（每天 75/75IU）治疗 3 天，接着接受 hCG（2000IU）治疗。hCG 后 36 小时，通过腹腔镜获取卵母细胞。对中期 II（MII）卵母细胞进行胞浆内单精子注射。将受精胚胎置于不同的培养条件和饲养细胞共培养中。通过最先进的阶段观察胚胎发育，包括囊胚形成。
卵母细胞回收数、受精率、多细胞胚胎率和囊胚率。
从 49 个周期中回收了 1924 个食蟹猴卵母细胞。外源性促性腺激素反应和随后的卵母细胞成熟存在明显的异质性。MII 卵母细胞的百分比在个体食蟹猴和刺激周期之间没有显著差异。近三分之二的 MII 卵母细胞通过 ICSI 成功受精。不同处理的胚胎囊胚率差异显著。在 P1/Blast、Quinn's Advantage 和 Sydney IVF 培养基中与饲养细胞共培养可产生更高的囊胚率。
我们测试了多种培养基和饲养细胞组合，以优化食蟹猴胚胎培养的条件，获得了类似于体外培养恒河猴胚胎的高囊胚率，但仍低于女性辅助生殖技术的囊胚率。
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