Beyond IUI, IVF and ICSI - New developments in - eshre 2013
Beyond IUI, IVF and ICSI - New developments in - eshre 2013
Beyond IUI, IVF and ICSI - New developments in - eshre 2013
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Case Study<br />
• Couple exhibit<strong>in</strong>g long-term <strong>in</strong>fertility (10+years) associated with high levels of DNA damage <strong>in</strong> the<br />
male germ l<strong>in</strong>e. Patient produced an oligozoospermic ejaculate conta<strong>in</strong><strong>in</strong>g 3.2 million<br />
spermatozoa/ml <strong>and</strong> an equivalent number (2.1 million/ml) of contam<strong>in</strong>at<strong>in</strong>g round cells.<br />
• Pre-separation:<br />
30% vitality<br />
18% motility<br />
DNA damage - 26% TUNEL positive, 41% SCSA DFI<br />
• Post-separation:<br />
62% vitality<br />
24% motility<br />
DNA damage - 14% TUNEL positive, 15% SCSA DFI<br />
• Intracytoplasmic sperm <strong>in</strong>jection (<strong>ICSI</strong>) was conducted us<strong>in</strong>g the electrophoretically isolated<br />
spermatozoa.<br />
• Oocytes were fertilized <strong>and</strong> normal blastocysts were generated after 5 days of culture. Transfer of<br />
two embryos was associated with the generation of a positive hCG signal followed by confirmation<br />
of a viable pregnancy by ultrasound.<br />
This is the first cl<strong>in</strong>ical report of a viable pregnancy follow<strong>in</strong>g the electrophoretic isolation of<br />
spermatozoa<br />
Conventional methodology<br />
Electrophoretic sperm separation<br />
Percentage (%)<br />
Embryo quality<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
2 PN Oocytes Cleavage Grade<br />
4-5 embryos<br />
Pregnancy rate<br />
L<br />
Prelim<strong>in</strong>ary Cl<strong>in</strong>ical trial<br />
Percentage P (%)<br />
Sperm quality<br />
80<br />
When all embryos are derived from one sperm preparation method<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
0<br />
Recovery Motility Concentration<br />
Conventional Electrophoretic<br />
-<br />
Embryo transfers 11 18<br />
Pregnancies 2 (18%) 6 (33%)<br />
Ratio of X <strong>and</strong> Y bear<strong>in</strong>g spermatozoa<br />
PCR amplicons of the amelogen<strong>in</strong> gene<br />
kb<br />
1.4<br />
1.2<br />
1.0<br />
0.8<br />
XX XY<br />
977 bp<br />
788 bp<br />
kb Increas<strong>in</strong>g ratio of male to female DNA<br />
1.4<br />
1.2<br />
1.0<br />
977 bp<br />
0.8<br />
788 bp<br />
L 1 2 3 4 5 6 7 8<br />
4 chamber system<br />
15 kDa restriction<br />
5 µm separation<br />
membranes<br />
membranes<br />
Anode (+ ve term<strong>in</strong>al)<br />
Y-bear<strong>in</strong>g sperm (%)<br />
80<br />
60<br />
40<br />
20<br />
0<br />
Cathode (- ve term<strong>in</strong>al)<br />
Cathode<br />
Anode<br />
Con S 1 2 3<br />
A<strong>in</strong>sworth et al, 2007<br />
Flem<strong>in</strong>g et al., 2008<br />
A<strong>in</strong>sworth et al, 2011<br />
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