Human fertilization with round and elongated spermatids |
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Authors: | Fishel S; Green S; Hunter A; Lisi F; Rinaldi L; Lisi R; McDermott H |
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Institution: | NURTURE (Nottingham University Research and Treatment Unit in Reproduction), Department of Obstetrics and Gynaecology, Queen's Medical Centre, UK. |
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Abstract: | 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.
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