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

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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 Page 26 of 127
Cathode (- ve terminal) Anode (+ ve terminal) kD 170 130 100 72 55 40 Mechanism – sialation high quality sperm SN NA binding (%) 100 80 60 40 20 Cathode Anode S 1 2 3 0 Washed Not washed Seminal plasma proteins 33 Cathode ** Anode ** ** S 1 2 3 Seminal plasma proteins co-migrate with sperm and suppress capacitation ** kD 71 40 30 Aminopeptidase B Fucosyltransferase 1 Prostatic acid phosphatase Unknown Preparation of spermatozoa for IVF Cathode (- ve terminal) Anode (+ ve terminal) Soerm bound/egg 35 30 25 20 15 10 *** 5 0 S ES Tyrosine phosphorylation Cathode Anode S 1 2 3 These cells can be used directly for IVF without washing ES S ES S Percoll Ainsworth et al, 2011 Ainsworth et al, 2011 Electrophoretic Sperm Separation – cassette design Buffer flow Asymmetrical - IUI Buffer flow Cathode (-ve terminal) Innoculation chamber Separation membrane Collection chamber Anode (+ve terminal) Symmetrical – General use Symmetrical - IVF and ICSI 15 kDa restriction 5 µm separation membranes membranes Anode (+ ve terminal) Cathode (- ve terminal) Page 27 of 127
Page 1: 28th Annual Meeting of the European
Page 5 and 6: Contents Course coordinators, cours
Page 7 and 8: Course coordinators Herman Tournaye
Page 9 and 10: Scientific programme Sperm selectio
Page 11 and 12: ESHRE SIG Andrology, Istanbul, Turk
Page 13 and 14: October 2001 - “A Physics Revelat
Page 15 and 16: Polydimethylsiloxan and Sperm Survi
Page 17 and 18: Microfluidic Sperm Isolation Reduce
Page 19 and 20: Micro-insemination Device 1. Remove
Page 21 and 22: Selecting spermatozoa; potential ro
Page 23 and 24: Fertilization Rate (%) 120 100 80 6
Page 25 and 26: Spermsep CS10 System Sperm Cell Sep
Page 27: TUNEL (%) 14% 90 80 70 60 50 40 30
Page 31 and 32: Hydrogen peroxide mediates DNA dama
Page 33 and 34: ESHRE 2012: Pre‐congress course 2
Page 35 and 36: Sperm morphology and ICSI The estab
Page 37 and 38: Motile Sperm Organellar Morphology
Page 39 and 40: Conclusions of the meta‐analysis
Page 41 and 42: Does the presence of sperm nuclear
Page 43 and 44: Basic sperm parameters Sperm DNA fr
Page 45 and 46: No relationship between chromosome
Page 47 and 48: Gynecology: Filippo Ubaldi Elena Ba
Page 49 and 50: Sperm selection: why? There is an i
Page 51 and 52: I- MACS -Annexine V for diagnostic
Page 53 and 54: Evaluation of HA-binding and sperm
Page 55 and 56: HA-bound sperm and morphology with
Page 57 and 58: III. HA and ICSI outcome: prospecti
Page 59 and 60: HA- clinical uses and limitations
Page 61 and 62: May the force be with you Using imm
Page 63 and 64: Sperm ultrastructure Centrosome cha
Page 65 and 66: Diagnostic semen analysis Three ba
Page 67 and 68: Ultrastructural defects Transmissio
Page 69 and 70: Necrozoospermia / low viability rat
Page 71 and 72: 1. Hypo-osmotic swelling test ICSI
Page 73 and 74: 4. Laser-assisted selection (LAISS)
Page 75 and 76: 5. Birefringence-polarisation micro
Page 77 and 78: Conclusions Absolute asthenozoospe
Page 79 and 80:
Definition of Azoospermia: Absence
Page 81 and 82:
Cases of aspermia Preventive measur
Page 83 and 84:
The probability crypto azoospermic
Page 85 and 86:
"The beauty and the beast" when the
Page 87 and 88:
Fertilization rate after ICSI 1- Oo
Page 89 and 90:
With (ICSI), zona pellucida penetra
Page 91 and 92:
Sibling oocytes from each patient w
Page 93 and 94:
The electron microscopic results in
Page 95 and 96:
Ionophore treatment for oocyte acti
Page 97 and 98:
The oocytes were suspended in 0.3M
Page 99 and 100:
Table 3. Pregnancy rates and outcom
Page 101 and 102:
References • Devroey P, Van Steir
Page 103 and 104:
Longing for a girl: Gender selectio
Page 105 and 106:
1,1 1 0,9 0,8 0,7 0,6 Sex ratio imb
Page 107 and 108:
PhD Thesis Michelle Duc, 1977 Retro
Page 109 and 110:
Reasons for wanting a girl at GC
Page 111 and 112:
Assessment of cervical mucus OOpaqu
Page 113 and 114:
Study design • A reference group
Page 115 and 116:
Partial data Reason n No post‐pre
Page 117 and 118:
Percentage P of girls 70 60 50 40 3
Page 119 and 120:
New aspects of this research • Fi
Page 121 and 122:
Mark your calendar for the upcoming
Page 123 and 124:
NOTES Page 121 of 127
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Beyond IUI, IVF and ICSI - New developments in - eshre 2013

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