t
test, and frequencies were compared using Fisher's exact
test. A
P
value of
<
.05 was considered signi
f
cant in all
circumstances.
RESULTS
We performed two- and three-color FISH analysis of sperm
chromosomes 13, 18, 21, X, and Y on ejaculated sperm
from 140 men who had RPL. Average sperm aneuploidy
was greater in sperm from men with RPL than in normozoo-
spermic controls (sex chromosome: 1.04% vs. 0.38%,
P
¼
.015; chromosome 18: 0.18% vs. 0.03%,
P
<
.001; chromo-
somes 18/21: 0.26% vs. 0.08%,
P
¼
.002) (
Table 1
). A higher
proportion of men with RPL had sperm aneuploidy (
>
2 SDs
above the mean aneuploidy for controls) for the sex chromo-
some compared with controls (53% vs. 3%,
P
<
.001). Interest-
ingly, there was no difference between the proportion of men
with RPL and aneuploidy and controls for either chromosome
18 (36% vs. 29%,
P
¼
.25) or chromosomes 13/21 (50% vs.
53%,
P
¼
.71).
A total of 40% of men with normal sperm density and
motility had increased sperm aneuploidy in both sex chromo-
somes and autosomes (
Table 2
) compared with controls. On
evaluating sperm aneuploidy in men with abnormal semen
parameters, we found that a larger proportion of men with
isolated low sperm density (
<
15
±
10
6
/mL) and low sperm
motility (
<
40%) had greater sperm sex chromosome aneu-
ploidy compared with men with normal sperm density and
motility (62% vs. 45%,
P
¼
.042) (
Table 2
). Similarly, there
was a greater proportion of men with abnormal strict
morphology (
<
4%) who also had sex chromosome aneu-
ploidy (57% vs. 28%,
P
¼
.04) compared with men with normal
strict morphology (
>
4%) (
Table 3
). As expected, there was
no association between abnormal DNA fragmentation
(
>
30%) and severity of aneuploidy or aneuploidy rates
(
Supplemental Table 1
, available online).
DISCUSSION
With the increased use of IVF/ICSI for male factor infertility, it
is important to identify reasons for failure. One of the greatest
challenges with ICSI is the identi
f
cation of
‘‘
normal sperm
’’
for micro-manipulation. Unfortunately, with current technol-
ogies, we can only identify sperm with grossly abnormal
morphology rather than detecting underlying genetic abnor-
malities, such as aneuploidy.
Controversy exists concerning safety of ICSI and whether
using genetically defective sperm will lead to abnormal fe-
tuses or IVF failure. Several studies have demonstrated an
increased frequency of genetic abnormalities in men with
TABLE 2
Prevalence of fragmentation, morphology, and abnormal FISH in men with normal/abnormal semen parameters.
Variable
Abnormal sperm
density and motility
Normal sperm
density and motility
P
value
Age (y)
38.0
²
6.6
37.4
²
5.4
.535
SM (%)
1.45
²
1.47
2.56
²
1.93
.001
Density (
±
10
6
/mL)
13.6
²
19.3
57.7
²
35.3
<
.001
Motility (%)
30.5
²
19.4
59.9
²
11.2
<
.001
Sex disomy (%)
0.01
²
0.01
0.10
²
0.80
.361
18 disomy (%)
0.23
²
0.53
0.14
²
0.34
.267
13/21 disomy (%)
0.36
²
0.91
0.18
²
0.29
.132
Men with sex chromosome disomy, % (n)
62.3 (40)
44.7 (34)
.042
Men with chromosome 18 disomy, % (n)
37.5 (24)
35.5 (27)
.861
Men with chromosomes 13/21 disomy, % (n)
56.3 (36)
44.7 (34)
.235
Note:
Values are mean
²
SD, unless otherwise noted. SM
¼
strict morphology.
Ramasamy. Sperm FISH and recurrent pregnancy loss. Fertil Steril 2015.
TABLE 3
Comparison of sperm aneuploidy in men with normal and abnormal strict morphology.
Variable
SM
<
4%
SM
>
4%
P
value
Age (y)
37.9
²
5.7
35.6
²
5.9
.149
SM (%)
1.46
²
1.10
5.56
²
0.84
<
.001
Density (
±
10
6
/mL)
34.4
²
32.5
66.9
²
45.8
<
.001
Motility (%)
45.0
²
21.0
60.6
²
13.8
.001
Sex disomy (%)
0.08
²
0.70
0.00
²
0.01
.276
18 disomy (%)
0.20
²
0.47
0.05
²
0.11
.005
13/21 disomy (%)
0.28
²
0.75
0.15
²
0.49
.479
Men with sex chromosome disomy, % (n)
56.6 (56)
27.8 (5)
.038
Men with chromosome 18 disomy, % (n)
40.5 (99)
16.7 (3)
.066
Men with chromosomes 13/21 disomy, % (n)
50.5 (50)
27.8 (5)
.122
Note:
Values are mean
²
SD, unless otherwise noted.
Ramasamy. Sperm FISH and recurrent pregnancy loss. Fertil Steril 2015.
908
VOL. 103 NO. 4 / APRIL 2015
ORIGINAL ARTICLE: ANDROLOGY