he future of cryopreservation in assisted reproduction _(by Handyside_Alan_2010)

1. ALPHA 2010
8th Biennial Conference
Budapest, Hungary
30th April - 2nd May, 2010
The future of cryopreservation
in assisted reproduction
Alan H Handyside
London Bridge Fertility, Gynaecology
And Genetics Centre
London
United Kingdom

2. SELECTION FREEZING
GENETIC SCREENING

3. Which
single
embryo to
select for
transfer?

4. Which
single
embryo to
select for
transfer?

5. Comparative Genomic
Hybridisation (CGH)

6. CONTROL TEST

7. CONTROL TEST
1.5
1.0
0.5

8. CONTROL TEST
1.5
1.0
0.5

9. CONTROL TEST
1.5
1.0
0.5

10. First or first and second polar
body biopsy on day 0 or day 1
Array comparative
genomic hybridisation
(CGH) following
whole genome
amplification

11. September 2nd 2009
New IVF test – Array
CGH – produces baby
Oliver, offering hope
to infertile
13 previous failed IVF
cycles
8/12 first polar bodies
aneuploid

12. 6.7 Mb intersitial duplication in 11q24.2g25 aascoiated with
infertility, minor dysmorphic features and normal
psyshomotor development
Gohring et al (2008) Eur J Med Genet Epub

13. Clinical application of comprehensive
chromosome screening at the blastocyst stage
• Zona drilling and blastocyst biopsy
• Vitrification using Crytop
• Chromosome copy number assessment by
metaphase comparative genomic
hybridisation (CGH) following whole genome
amplification
Schoolcraft et al (2009) Fertility and Sterility in press

14. • 45 patients (mean maternal age 37.7 yrs)
• 287 blastocysts (6.4 per cycle)
• 93.7% CGH successful
• 138 (51.3%) blastocysts aneuploid
• 100% survival following
biopsy/vitirifcation/thawing
• 82.2% pregnancy rate per cycle (mean 2.0
blastocysts transferred)
• 68.9% FHB/ blastocyst transferred
• 55% multiple pregnancies

15. Single nucleotide
polymorphism genotyping and
karyomapping
Handyside et al (2009) J Med Genet

16. Single nucleotide
polymorphisms (SNPs)
• 10 million SNPs across
human genome
• Many biallelic (AA, AB, BB)
• Major contribution to
genetic diversity, inherited
disease and variants
associated with common
multifactorial conditions

17. ♂ ♀
A A A B
A A A A
B B B A
A A B A
A B B B
B A A A

18. A ♂ ♀
Karyomapping combines
genome wide linkage based
detection of single gene defects
(A) with chromosomal
aneuploidy including
monosomy/deletions (B) and
trisomies involving inheritance of
two different meiotic B
chromosomes from one parent
(D). Chromosome duplication is
not detected (C).
C D

19. Family 2
Preimplantation genetic diagnosis
for cystic fibrosis
Whole genome amplification by
isothermal multiple displacement
amplification of 2-10 cells in each
biopsy/embryo

20. Improved cryopreservation methods
? Improved implantation rates in unstimulated cycles
? Improved birth weights and health
Highly resolution 24 chromosome screening
Less invasive polar body or blastocyst biopsy
Maximise live birth rates per cycle
Avoid multiple pregnancies
Reduce pregnancy loss and miscarriage
Minimise FETs