Beyond IUI, IVF and ICSI - New developments in - eshre 2013

Case Study
• Couple exhibiting long-term infertility (10+years) associated with high levels of DNA damage in the
male germ line. Patient produced an oligozoospermic ejaculate containing 3.2 million
spermatozoa/ml and an equivalent number (2.1 million/ml) of contaminating round cells.
• Pre-separation:
30% vitality
18% motility
DNA damage - 26% TUNEL positive, 41% SCSA DFI
• Post-separation:
62% vitality
24% motility
DNA damage - 14% TUNEL positive, 15% SCSA DFI
• Intracytoplasmic sperm injection (ICSI) was conducted using the electrophoretically isolated
spermatozoa.
• Oocytes were fertilized and normal blastocysts were generated after 5 days of culture. Transfer of
two embryos was associated with the generation of a positive hCG signal followed by confirmation
of a viable pregnancy by ultrasound.
This is the first clinical report of a viable pregnancy following the electrophoretic isolation of
spermatozoa
Conventional methodology
Electrophoretic sperm separation
Percentage (%)
Embryo quality
120
100
80
60
40
20
0
2 PN Oocytes Cleavage Grade
4-5 embryos
Pregnancy rate
L
Preliminary Clinical trial
Percentage P (%)
Sperm quality
80
When all embryos are derived from one sperm preparation method
70
60
50
40
30
20
10
0
0
Recovery Motility Concentration
Conventional Electrophoretic
-
Embryo transfers 11 18
Pregnancies 2 (18%) 6 (33%)
Ratio of X and Y bearing spermatozoa
PCR amplicons of the amelogenin gene
kb
1.4
1.2
1.0
0.8
XX XY
977 bp
788 bp
kb Increasing ratio of male to female DNA
1.4
1.2
1.0
977 bp
0.8
788 bp
L 1 2 3 4 5 6 7 8
4 chamber system
15 kDa restriction
5 µm separation
membranes
membranes
Anode (+ ve terminal)
Y-bearing sperm (%)
80
60
40
20
0
Cathode (- ve terminal)
Cathode
Anode
Con S 1 2 3
Ainsworth et al, 2007
Fleming et al., 2008
Ainsworth et al, 2011
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