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853856, 2000
Detection of chromosomal abnormalities by fluorescent
in-situ hybridization in immotile viable spermatozoa
determined by hypo-osmotic sperm swelling test*
Hulusi Bulent Zeyneloglu1,3, Volkan Baltaci2,
immature forms of sperm cells from the testis may be obtained
Sevinc Ege1, Ali Haberal1 and S.Batioglu1
with varying success.
Poor or absent fertilization rates are obtained in cases of
1Department of Obstetrics/Gynaecology, and 2Department of
ICSI using immotile ejaculated spermatozoa of unknown
Medical Genetics, Baskent University, Ankara, Turkey
viability (Nagy et al. , 1995). Total immotility may be caused
3To whom correspondence should be addressed at: Mesa Koru
by necrozoospermia, ultrastructural defects or metabolic immaSitesi Manolya Blok 24, Cayyolu, 0630, Turkey
turity of the spermatozoa.
If randomly selected immotile spermatozoa are used for
In cases of total necrozoospermia in the ejaculate, motile
intracytoplasmic sperm injection (ICSI), pregnancy rates
spermatozoa can sometimes be recovered from a testicular
are significantly decreased. The hypo-osmotic swelling test
biopsy (Tournaye et al. , 1996). However, when selecting
(HOST) is the only method available to detect the viable,
spermatozoa to inject into an oocyte, it is always preferable
but immotile spermatozoa for ICSI. However, evidence is
to use the one that has shown some sign of motility and with
still lacking for the chromosomal abnormalities for the
best morphology available.
normal-looking, but immotile spermatozoa positive for
The hypo-osmotic swelling test (HOST) or cell staining
HOST. Sperm samples from 20 infertile men with normal
methods are often used to identify sperm viability. The mechanchromosomal constitution were obtained. After Percoll
isms of HOST and eosin-Y staining are mainly related to the
separation, morphologically normal but immotile spermafunctional integrity of sperm membrane and normal functional
tozoa were transported individually into HOST solution
activity of human spermatozoa (Jeyendran et al. , 1984). Since
for 1 min using micropipettes. Cells that showed tail curling
there is no available cell-staining technique that does not
with swelling in HOST were then transferred back into
impair the viability of the cell for future clinical use, HOST
human tubal fluid solution to allow reversal of swelling.
is the most useful method of selection for the viable spermatoThese sperm cells were fixed and processed for the multi-
held conjointly with the 54th Annual Meeting of the American Society
immotile spermatozoa were transported individually into HOST
for Reproduction, October 49, 1998, San Francisco, CA, USA.
solution (Hypo-10, Scandinavian Scientific, Gothenburg, Sweden)
European Society of Human Reproduction and Embryology
853
H.B.Zeyneloglu et al.
Figure 2. Photographs of immotile spermatozoa after FISH analysis
with centromeric probes for the chromosomes 18 (blue), X (green)
Figure 1. The spermatozoa incubated in hypo-osmotic solution, the
and Y (orange) in a three-colour hybridization protocol. All probes
HOST () spermatozoa demonstrate curling and swelling of the
were directly labelled with fluorochromes and obtained
tails. Scale bar 5 m.
commercially. Scale bar 5 m.
using micropipettes (Hunter Scientific, Saffron Walden, Essex UK)
of a microinjection device (Nikon, Narishige, Japan); if the final
injection is not related to any of the three basic sperm parameters. Hum.
Reprod. , 10, 11231129.
of the chromosomal abnormalities. In conclusion, immotile
Neugebauer, D.C., Neuwinger, J., Jockenhovel, F. et al. (1990) 9 0
spermatozoa from men with normal karyotype in the absence
axoneme in spermatozoa and some nasal cilia of a patient with totally
of microdeletions of the Y chromosome may be safely used
immotile spermatozoa associated with thickened sheath and short midpiece.
Hum. Reprod. , 5, 981986.
for ICSI purposes.
Olmedo, S.B., Nodar, F., Chillik, C. et al. (1997) Successful intracytoplasmic sperm
injection with spermatozoa from a patient with dysplasia of the
fibrous sheath and chronic respiratory disease. Hum. Reprod. , 12, 14971499.
Palermo, G., Joris, H., Devroey, P. et al. (1992) Pregnancies after
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