Human Reproduction, Vol 13, 639-645, Copyright © 1998 by Oxford University Press
I Aslam, A Robins, K Dowell and S Fishel
The success of spermatid microinjection has generated many concerns. In
particular, there is a lack of appropriate methodology for the isolation of
large homogeneous populations of spermatids, with minimum loss of
viability, from the testicular tissue of azoospermic men. Here we have
compared two different isolation methods -- velocity sedimentation under
unit gravity (VSUG) combined with discontinuous Percoll centrifugation
(DPC), and separation with fluorescent-activated cell sorter (FACS) using
light in the visible range -- to determine the most suitable method for the
isolation of spermatids. Total mixed cell count/gram of testicular
parenchyma was significantly higher in obstructive azoospermic men compared
with non-obstructive azoospermic men (P < 0.001). The results of the
comparison showed that in obstructive azoospermic patients the difference
in the yields of primary spermatocytes produced by the two techniques was
not significant, but for round and elongating spermatids the FACS
separation proved to be the better method (P < 0.001). Similarly, in
non-obstructive azoospermic patients, FACS separation proved to be
superior, giving increased yields of primary spermatocytes and round and
elongating spermatids compared with VSUG combined with DPC method (P <
0.001). More than 99 % of the separated cells retained their viability
after FACS separation. As large homogeneous populations of viable
spermatids can be separated with FACS in a relatively short period of time,
FACS separation is the most suitable method for the isolation of spermatids
from testicular biopsy tissue.
ARTICLES
Isolation, purification and assessment of viability of spermatogenic cells from testicular biopsies of azoospermic men
CARE, The Park Hospital, Arnold, Nottingham, UK.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Coskun, A. Tbakhi, K. A. Jaroudi, M. Uzumcu, T. A. Merdad, and K. A. Al-Hussein Flow cytometric ploidy analysis of testicular biopsies from sperm-negative wet preparations Hum. Reprod., April 1, 2002; 17(4): 977 - 983. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Johnson, C. Staub, W. B. Neaves, and R. Yanagimachi Live human germ cells in the context of their spermatogenic stages Hum. Reprod., August 1, 2001; 16(8): 1575 - 1582. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Aslam, S. Fishel, H. Moore, K. Dowell, and S. Thornton Fertility preservation of boys undergoing anti-cancer therapy: a review of the existing situation and prospects for the future: Opinion Hum. Reprod., October 1, 2000; 15(10): 2154 - 2159. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Prapas, A. Chatziparasidou, P. Vanderzwalmen, M. Nijs, N. Prapas, B. Lejeune, G. Vlassis, and R. Schoysman Spermatid injection: Reconsidering spermatid injection Hum. Reprod., September 1, 1999; 14(9): 2186 - 2188. [Full Text] [PDF]
|
|
|
|
|
|
P. Sutovsky, J. Ramalho-Santos, R. D. Moreno, R. Oko, L. Hewitson, and G. Schatten On-stage selection of single round spermatids using a vital, mitochondrion-specific fluorescent probe MitoTrackerTM and high resolution differential interference contrast microscopy Hum. Reprod., September 1, 1999; 14(9): 2301 - 2312. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Aslam and S. Fishel Evaluation of the fertilization potential of freshly isolated, in-vitro cultured and cryopreserved human spermatids by injection into hamster oocytes Hum. Reprod., June 1, 1999; 14(6): 1528 - 1533. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Cremades, R. Bernabeu, A. Barros, and M. Sousa In-vitro maturation of round spermatids using co-culture on Vero cells Hum. Reprod., May 1, 1999; 14(5): 1287 - 1293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gandini, A. Lenzi, F. Lombardo, R. Pacifici, and F. Dondero Immature germ cell separation using a modified discontinuous Percoll gradient technique in human semen Hum. Reprod., April 1, 1999; 14(4): 1022 - 1027. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Ziyyat, B. Lassalle, J. Testart, P. Briot, E. Amar, C. Finaz, and A. Lefevre Flow cytometry isolation and reverse transcriptase–polymerase chain reaction characterization of human round spermatids in infertile patients Hum. Reprod., February 1, 1999; 14(2): 379 - 387. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Lassalle, A. Ziyyat, J. Testart, C. Finaz, and A. Lefevre Flow cytometric method to isolate round spermatids from mouse testis Hum. Reprod., February 1, 1999; 14(2): 388 - 394. [Abstract] [Full Text] [PDF]
|
|