Study question Does human recombinant hyaluronidase (Cumulase®) improve fertilization and blastocyst formation rates in sibling human oocytes compared to bovine-derived hyaluronidase? Summary answer The use of Cumulase® for oocyte denudation does not appear to affect fertilization or usable blastocyst formation rate in patients undergoing intracytoplasmic sperm injection (ICSI). What is known already Bovine-derived hyaluronidase has been widely employed to denude the cumulus-oocyte complexes (COCs) prior to ICSI or oocyte vitrification. Nonetheless, the recent concern for its safety, due to its animal origin, has led to the development of alternatives ascertaining the enzyme’s purity. It has been reported that human recombinant hyaluronidase is not equally effective as its bovine-derived analogue, leading to a longer time required for complete denudation coupled by the subsequent toxicity risk for oocytes. This may result to lower fertilization and blastocyst formation rates. Although there are some studies on IVF outcomes with recombinant hyaluronidase, its use remains rather limited. Study design, size, duration This randomized double-blinded clinical trial was conducted at Citmer Reproductive Medicine, in Puebla and Monterrey, Mexico from February to December 2021. The impact of denudation on sibling oocytes from 103 patients employing bovine-derived hyaluronidase (InVitroCare®) or human recombinant hyaluronidase (ICSI Cumulase® (rHuPH20), CooperSurgical®) was evaluated. A total of 1237 oocytes were assigned as follows: The Hyaluronidase Group included 626 oocytes, while the Cumulase® Group included 611 oocytes. Preimplantation Genetic Testing was performed in 18 cycles. Participants/materials, setting, methods After retrieval, COCs were incubated for 2-4 h and placed in microdropets with 10 IU/mL bovine-derived hyaluronidase or Cumulase® in culture media GlobalTotal® (LifeGlobal®) for granulosa cells removal. COCs were gently pipetted up and down employing 20 μL and 150 μm tips. Metaphase II oocytes were injected and incubated for 17-20 hours in 8% CO2, 20% O2, 37°C. Normally fertilized zygotes were cultured to blastocyst stage and embryo development was evaluated. Seventy-nine blastocysts were biopsied. Main results and the role of chance The mean age of the patients was 32.12±1.11 years old. A total of 506 and 493 metaphase II oocytes from Groups 1 and 2 respectively were subjected to ICSI. Normal fertilization rates for oocytes treated with Hyaluronidase and Cumulase® were 71.34% (n = 361) and 72.41% (n = 357) respectively, with no statistically significant differences between the two groups (p = 0.35). Usable blastocysts rate (selected for embryo transfer or blastocyst cryopreservation) did not differ with any statistical difference between the bovine-derived hyaluronidase (n = 170, 47.09%) and the Cumulase® treated oocytes (n = 158, 44.25%), (p = 0.44). Preimplantation genetic testing for aneuploidies (PGT-A) was performed in 18 cycles. Seventy-nine blastocysts were biopsied and tested. Fourty-seven embryos from the bovine-derived hyaluronidase group were biopsied, presenting with a euploidy rate of 51.06% (n = 24), compared to 32 embryos from the recombinant hyaluronidase group with a euploidy rate of 50% (n = 16). Results indicated no statistically significant difference (p = 0.92). These data support that human recombinant hyaluronidase presents with similar efficacy to bovine-derived hyaluronidase. in terms of normal fertilization, embryo development as well as euploidy rates. Limitations, reasons for caution The small sample size regarding the euploidy outcome stands as a limitation of the study. Further studies should focus on the effect of recombinant hyaluronidase on genetic anomalies on blastocysts. Wider implications of the findings Fertilization, usable blastocyst and euploidy rates did not differ when employing bovine-derived or human recombinant hyaluronidase, rendering the latter a safe and efficient alternative. However, it is important to assess long-term effects, as only a limited number of studies report on perinatal and neonatal outcomes Trial registration number 25-12-21MTYPUECIT

中文翻译：

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P-148 使用人重组透明质酸酶的剥脱术不影响接受 ART 患者同胞卵母细胞的受精和囊胚形成率：一项前瞻性随机研究

研究问题 与源自牛的透明质酸酶相比，人类重组透明质酸酶 (Cumulase®) 是否能提高同胞人类卵母细胞的受精率和囊胚形成率？总结回答 使用 Cumulase® 进行卵母细胞剥脱似乎不会影响接受胞浆内单精子注射 (ICSI) 的患者的受精或可用囊胚形成率。已知的牛源性透明质酸酶已广泛用于在 ICSI 或卵母细胞玻璃化之前剥去卵丘-卵母细胞复合物 (COC)。尽管如此，由于其动物来源，最近对其安全性的担忧已导致开发确定酶纯度的替代品。据报道，人重组透明质酸酶不如其牛衍生的类似物有效，导致完全剥脱所需的时间更长，加上随后对卵母细胞的毒性风险。这可能导致受精率和胚泡形成率降低。尽管有一些关于重组透明质酸酶体外受精结果的研究，但其使用仍然相当有限。研究设计、规模、持续时间这项随机双盲临床试验于 2021 年 2 月至 2021 年 12 月在墨西哥普埃布拉和蒙特雷的 Citmer Reproductive Medicine 进行。剥蚀对 103 名使用牛源性透明质酸酶 (InVitroCare®) 患者的同胞卵母细胞的影响) 或人重组透明质酸酶 (ICSI Cumulase® (rHuPH20), CooperSurgical®) 进行了评估。共有 1237 个卵母细胞分配如下：透明质酸酶组包括 626 个卵母细胞，而 Cumulase® 组包括 611 个卵母细胞。植入前基因检测在 18 个周期内进行。参与者/材料、设置、方法 取回后，将 COC 孵育 2-4 小时，然后将其置于培养基 GlobalTotal® (LifeGlobal®) 中含有 10 IU/mL 牛源性透明质酸酶或 Cumulase® 的微滴中，以去除颗粒细胞。使用 20 μL 和 150 μm 吸头轻轻地上下移取 COC。注射中期 II 卵母细胞并在 8% CO2、20% O2、37°C​​ 中孵育 17-20 小时。将正常受精的受精卵培养至囊胚期并评价胚胎发育。对 79 个囊胚进行了活检。主要结果及机会的作用 患者平均年龄为32.12±1.11岁。来自第 1 组和第 2 组的总共 506 和 493 个中期 II 卵母细胞分别接受 ICSI。用透明质酸酶和 Cumulase® 处理的卵母细胞的正常受精率分别为 71.34% (n = 361) 和 72.41% (n = 357)，两组之间无统计学差异 (p = 0.35)。可用囊胚率（选择用于胚胎移植或囊胚冷冻保存）在牛源性透明质酸酶（n = 170, 47.09%）和 Cumulase® 处理的卵母细胞（n = 158, 44.25%）之间没有任何统计学差异，（p = 0.44)。非整倍体植入前基因检测 (PGT-A) 在 18 个周期内进行。对七十九个囊胚进行了活检和测试。来自牛源性透明质酸酶组的 47 个胚胎进行了活检，整倍性率为 51.06%（n = 24），而重组透明质酸酶组的 32 个胚胎整倍性率为 50%（n = 16）。结果表明没有统计学上的显着差异（p = 0.92）。这些数据支持人重组透明质酸酶具有与牛衍生透明质酸酶相似的功效。在正常受精、胚胎发育以及整倍体率方面。局限性，谨慎的原因 关于整倍体结果的小样本量是该研究的局限性。进一步的研究应集中在重组透明质酸酶对胚泡遗传异常的影响。研究结果的更广泛意义 受精率、可用囊胚率和整倍体率在使用牛衍生或人类重组透明质酸酶时没有差异，这使得后者成为一种安全有效的替代方案。然而，评估长期影响很重要，