Non invasive emNon-invasive em
Barry Behr, y ,Prof
Director, IVF/ARDirector, IVF/ARCo-Director, R
Dept ODept OStanford Universi
mbryo Evaluationmbryo Evaluation
Ph.D., HCLD,fessorRT LaboratoriesRT LaboratoriesEI/IVF Program
Ob/GynOb/Gynity Medical Center
Disclo
I am a founder of AuxI am a founder of BlasI am a founder of Blas
osure
xogynstogen/IviGenstogen/IviGen
Out
BackgroundHistoric perspectiveHistoric perspectiveCurrent approachesF t t h l iFuture technologies
tline
What’s ImThe Patie
stimulatio
What culture system
stimulatio
What media Conditions/Exp
IVF outcome
mportant ?ent
on
Which stage
on
Which embryoertise
e
IVF Field AwaitingIVF Field Awaiting
“Young fertilitYoung fertilitfigure out wmake a babThat will reThat will remultiples bsuccess ratwhich curresurprisinglyhis team wDr. Howard Georgeanna Jones,
First IVF Baby and First IVF Clinic in the US
Epstein, Randi Hutter. “Pioneer Reflects o
g a Breakthroughg a Breakthrough
ty investigators today shouldty investigators today should which one embryo is likely to by rather than transfer several.educe costs, the number ofeduce costs, the number of births and significantly increase es of in vitro fertilization, ently hover around 30%‐‐y close to the 28% success rate as seeing in the 1980s.”
5
on Future of Reproductive Medicine.”, 22,March,2010
Risks of Multiple Em
Short‐ and long‐term risks to offsprin– Increased chance of miscarriage
Lo birth eight (LBW) and pre term b– Low birth weight (LBW) and pre‐term b
– 80% of infant mortalities result from 8%
Risks to the mother– Pre‐eclampsia / hypertensive disorder
– Pregnancy‐induced diabetes
– Miscarriage and other prenatal complic– Miscarriage and other prenatal complic
Selective fetal reduction
mbryos Transferred
ng
irth occ r 7 and 5 resp (Barker H pothesis)irth occur 7x and 5x, resp. (Barker Hypothesis)
% LBW, 13% pre‐term births
cations often requiring hospitalizationcations often requiring hospitalization
Whichh one?
Traditional
The Balancing Act !The Balancing Act !
L k f
Culture induced s
Lack of predictability
vs.
Low Implantation
Approach
Attrition in
stress
Attrition in culture
Hi h I l t tiHigh Implantation
Problems with Em
SubjectivePoor standardizationPoor standardizationTiming dependentC i dCan compromise devePre vs Post genomic aPaternal contribution a
mbryo Assessment
l telopmentactivationassessment
What you see is noWhat you see is noge
ot always what youot always what you et!
But…morphologyy isn’t everything!
Current EmbryCurrent EmbryEmbryos are evalua
morphological assess
Step 2: Occasional monitor
Step 1: Embryos develop in Occasional monitor
and Day 3 selectioEmbryos develop in
incubator
yo Assessmentyo Assessmentated based on simplesment on days 1 and 3
Non-ViableViable Viable
Viablering
Non-ViableNon-Viable
ring on
The dilem
BOTH RESULTED
mma……..
IN A SINGLETON P.Nagy
Human Embryoo Development
Important facto
Incubation chamber pH levelspO2 levelsOil vs. open culturepVolume of mediumOocyte/Embryo densityOocyte/Embryo densityMode of fertilization
ors to considerThe time of gamete co-incubationAssisted hatchingPGDEmbryo transfer techniqueOvarian stimulation protocols
Ultimat
Achieve a healthy singTransfer fewer embryoTransfer fewer embryo
te Goal
gleton pregnancyos (1 or 2)os (1 or 2)
New TechNew Techhnologieshnologies
What can
What do you need to b– Good genesGood genes– Good metabolism
n we do?
be healthy?
Embryo SelectEmbryo SelectMost technologies lack clinical trials
improved blastocyst predicimproved blastocyst predic
GenChemistry
Approach
• Full Karyotypeimplantation GScreening (PG
y• Metabolomic profiling
• Amino Acid uptake
• Cummulus Cell analysis
Limitation
•Mosaicism?
• Experts disageffectiveness
• Invasive proc
• Technology adoption challenge
• Doesn’t fit in current workflow • Invasive proc
ion Landscapeion Landscapes. No results exist to demonstrate ction or pregnancy ratesction or pregnancy rates.
etics Imaginge ‐ Pre‐Genetic GS)
g g• Unisense/Primo Vision– Time lapse imaging of
embryos• Olympus/Sanyo/Nikon/AstecOlympus/Sanyo/Nikon/Astec– Instrumentation
gree on s?
cedure
•No predictive parameters (yet)
•No prospective studies to show efficacy
• Lack of human data emergingcedure • Lack of human data , emerging
Amino Acid Upta
Immediate goal: 30%success rates.Ultimate goal - 50% iEmbryo Transfer ratesytwo or more embryos
ake: Novocellus
% increase in IVF
ncrease in Single s to match those with replaced
Novoc
Need to use proprietarCulture in micro volumCulture in micro volum
cellus
ry mediamesmes
“Omics” eOmics e
Economics GenomicsGenomicsProteomics
eraera……….
The “oThe o
Functiona
omics”omics
al Phenotype
Proteo
Current approaches:– Not user friendlyNot user friendly– Not rapid– Not high through putNot high through put
omics
Proteo
Most proteins are NOTProteins used for interProteins used for inter
omics
T secretedrnal processesrnal processes
MetaboMetabo
Molecular MetabMetab
olomicsolomics
Biometricsbolonbolon
Biomarker SpectraBiomarker Spectra(by N(by N(by N(by N
0 08
ance
0 06
0.08
Heme
Abs
orba
0.04
0.06
R-O
Heme
Rel
ativ
e A
0.02
0.04
CH
R
600 00 8000600 700 800
Wavelen
al Signatures al Signatures NIR)NIR)NIR)NIR)
OHOH
NH
OH
0 900 10000 900 1000ngth (nm)
Human Embryo TiPaper Published Samples
Payne et al. Hum Reproduction 1997 50
2PN • ObsHum Reproduction 2PN
Lemmen et al.RBM Online 2008 102
2PN oocytes
• Rep• Cor
preg
Mio et al. Am J Obstet Gyn 2008 286
oocytes • Obs• Rep
Wong et al 242
• Idenby DWong et al.
Nature Biotechnology 2010 2422PN
y• Dem
exp• Dev
Pribenszky et al. RBM Online 2010 5
2PN • RepRBM Online 2010 2PN p
Meseguer et al. Hum Reprod 2011 247
2PN • Eva
Hashimoto et al. 2012 80 EHashimoto et al.Fert Steril 2012 80
2PN • Eva
Swann et. al.Fertil steril 2012 10 oocytes
or zygotes • Cor
ime-lapse StudiesConclusions
served details of the fertilization process to 20 hrs
ported PN appearance & disappearancerelated synchrony in nuclei appearance after 1st cleavage with gnancy success
served details of the fertilization processported two ICMs - monozygotic twins
ntified cell cycle parameters that predict blastocyst formationDay 2ymonstrated that parameters correlate to embryo gene pression dataveloped cell tracking software
ported a live birthp
aluated cell cycle parameters to implantation
l t d ll l t f bl t t litaluated cell cycle parameters for blastocyst quality
related cytoplasmic movements with Ca2+ oscillations.
Imaging Systemg g yUnisense EmbryoScope Pri
Olympus Niko
ms (currently available in the USA)mo Vision
n Biostation Astec real time monitoring system
what is the embwhat is the embi.e. non–invasive, qua
bryo equivalent?bryo equivalent?antitative assessment
SumSumMany opportunities to interrogate emb
Time laps imaging offers a lot of prom
Parameters of the first three mitotic diindicate success to blastocyst (>93% success/failure inherited (maternal).
Defects in underlying molecular prograbehavior.
Should be able to be measured other
Improved diagnostics = (Early transferImproved diagnostics (Early transferoutcomes and increased success.
marymarybryos.
mise. Prospective trials emerging.
visions prior to embryonic activation specificity and sensitivity); suggesting
ams underlie aberrant blastomere
ways?
r) fewer embryos reduced adverser), fewer embryos, reduced adverse
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