Vanderzwalmen P, Zech H, Birkenfeld A, Yemini M, Bertin G, Lejeune B, Nijs M, Segal L, Stecher A, Vandamme B, van Roosendaal E, Schoysman R
Schoysman Infertility Management Foundation, Vaartsraat, Vilvoorde, Belgium.
Hum Reprod. 1997 Jun;12(6):1203-13. doi: 10.1093/humrep/12.6.1203.
Spermatid microinjection into oocytes has proven to be a successful assisted reproduction procedure in the animal model and in the human species, since in the latter a few full-term pregnancies were actually obtained. Patients entering our spermatid injection study included those with a total absence of spermatozoa in the testicular tissue notwithstanding previous positive biopsies (n = 29): an obstructive problem (n = 3), secretory azoospermia (n = 26), and those with total arrest at the spermatogenesis level in previous explorative biopsies (n = 15). In the latter group, absence of spermatids was recorded in four cases. Mature, elongated, elongating and round spermatids (ROS) were injected in respectively 3, 2, 3, and 32 attempts. A total of 260 metaphase II oocytes were injected with ROS, 36 oocytes with spermatids at other stages of maturity. The rates of oocytes showing two pronuclei (2PN) and two polar bodies reached 22% and 64% respectively after injection of round or elongated-mature spermatids. The fertilization rate after ROS injection was influenced by the percentage of spermatozoa observed in a previous biopsy. Patients with a positive preliminary biopsy had significantly more 2PN (33%) when compared to those with a severe spermatogenic dysfunction and in whom no spermatozoa were found (only 11%) (P < 0.05). Incubation of oocytes in calcium ionophore after ROS injection had a positive effect on the rate of 2PN formation (36 versus 16%). Ninety per cent of all the normally fertilized oocytes cleaved. The percentage of grade A and B embryos depended on the type of injected cells: 12% after ROS and 30% with the other types of haploid cells. A total of 39 transfers resulted in five pregnancies: three full term with healthy babies delivered (one after ROS injection, and two after injection of an elongating and a mature spermatid), one 4 months ongoing (after elongating spermatid injection) and one miscarriage at 4 weeks (after elongated cell injection). Compared to our conventional intracytoplasmic sperm injection-testicular sperm extraction (ICSI-TESE) programme, the implantation rate after ROS injection was very low (5.5 versus 10.5%).
在动物模型和人类中，将精子细胞显微注射到卵母细胞已被证明是一种成功的辅助生殖程序，因为在人类中实际上已获得了一些足月妊娠。进入我们精子细胞注射研究的患者包括那些尽管之前活检呈阳性但睾丸组织中完全没有精子的患者（n = 29）：梗阻性问题（n = 3）、分泌性无精子症（n = 26），以及那些在之前的探索性活检中精子发生水平完全停滞的患者（n = 15）。在后者组中，有4例记录为没有精子细胞。分别对成熟、伸长、正在伸长和圆形精子细胞（ROS）进行了3次、2次、3次和32次注射尝试。总共向260个中期II卵母细胞注射了ROS，向36个卵母细胞注射了其他成熟阶段的精子细胞。注射圆形或伸长成熟精子细胞后，显示双原核（2PN）和两个极体的卵母细胞比例分别达到22%和64%。ROS注射后的受精率受之前活检中观察到的精子百分比影响。初步活检呈阳性的患者与严重精子发生功能障碍且未发现精子的患者相比，有显著更多的2PN（33%对仅11%）（P < 0.05）。ROS注射后用钙离子载体孵育卵母细胞对2PN形成率有积极影响（36%对16%）。所有正常受精的卵母细胞中有90%发生了分裂。A和B级胚胎的百分比取决于注射细胞的类型：ROS注射后为12%，其他类型单倍体细胞注射后为30%。总共进行了39次移植，获得了5次妊娠：3次足月分娩出健康婴儿（1次在ROS注射后，2次在注射伸长和成熟精子细胞后），1次妊娠持续4个月（在注射伸长精子细胞后），1次在4周时流产（在注射伸长细胞后）。与我们传统的卵胞浆内单精子注射-睾丸精子提取（ICSI-TESE）方案相比，ROS注射后的着床率非常低（5.5%对10.5%）。
