Fishel S, Green S, Hunter A, Lisi F, Rinaldi L, Lisi R, McDermott H
NURTURE (Nottingham University Research and Treatment Unit in Reproduction), Department of Obstetrics and Gynaecology, Queen's Medical Centre, UK.
Hum Reprod. 1997 Feb;12(2):336-40. doi: 10.1093/humrep/12.2.336.
Human spermatids from ejaculate and testicular tissue have been utilized for evaluating human fertilization by intracytoplasmic sperm injection (ICSI) and, where possible, compared with spermatozoa utilizing sibling oocytes. Round and elongated spermatids obtained from ejaculates were either prepared through Percoll gradients or isolated and washed individually using subzonal insemination needles (SUZI; 10-14 microm internal diameter). Seminiferous tubules obtained after biopsy were placed into HEPES-buffered Earle's medium and dissected using 21-gauge needles. Spermatogenic cells and spermatozoa were isolated and washed individually using SUZI needles. Spermatozoa were subsequently injected into the ooplasm using 5 microm (internal diameter) ICSI needles, whereas 8-9 microm (internal diameter) needles were used for spermatid injection. Only metaphase II oocytes (n = 207) were injected: 64 with round spermatids, 92 with elongated spermatids and 51 with spermatozoa; the fertilization rate was 30, 24 and 67% respectively. There was a significant (P < 0.001) increase in the fertilization rate using spermatozoa compared with spermatids. The fertilization rate was not different between round and elongated spermatids, although the fertilization rates for round and elongated spermatids in the ejaculate were 33 and 18% respectively, compared with 22 and 38% respectively when testicular spermatids were utilized. In three patients sibling oocytes were used to compare round and elongated spermatids found in the ejaculate with spermatozoa extracted from seminiferous tubules. The fertilization rate was 24% for spermatids and 79% for testicular spermatozoa. This result suggests that, should only spermatids be available in the ejaculate, a testicular biopsy in the hope of obtaining testicular spermatozoa would be worth while.
来自射精和睾丸组织的人类精子细胞已被用于通过卵胞浆内单精子注射（ICSI）评估人类受精情况，并在可能的情况下，与使用同胞卵母细胞的精子进行比较。从射精中获得的圆形和延长型精子细胞，要么通过Percoll梯度制备，要么使用 zona 内授精针（SUZI；内径10 - 14微米）单独分离和洗涤。活检后获得的曲细精管被放入HEPES缓冲的Earle培养基中，并用21号针进行解剖。生精细胞和精子使用SUZI针单独分离和洗涤。随后使用内径5微米的ICSI针将精子注射到卵质中，而内径8 - 9微米的针用于精子细胞注射。仅注射了中期II卵母细胞（n = 207）：64个注射圆形精子细胞，92个注射延长型精子细胞，51个注射精子；受精率分别为30%、24%和67%。与精子细胞相比，使用精子时受精率显著提高（P < 0.001）。圆形和延长型精子细胞的受精率没有差异，尽管射精中圆形和延长型精子细胞的受精率分别为33%和18%，而使用睾丸精子细胞时分别为22%和38%。在三名患者中，使用同胞卵母细胞比较射精中发现的圆形和延长型精子细胞与从曲细精管中提取的精子。精子细胞的受精率为24%，睾丸精子的受精率为79%。这一结果表明，如果射精中仅存在精子细胞，为获取睾丸精子而进行睾丸活检是值得的。
