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Dr. Marcos MeseguerMarcos.meseguer@ivi.es

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To achieve high implantation rates we need “healthy embryos”

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…the oocyte is a very large cell and the cytoplasm is perhaps more important than it’s nucleus…

…the oocyte is a very large cell and the cytoplasm is perhaps more important than it’s nucleus…

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We must consider redefining what are “healthy embryos”

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To identify healthy embryos with healthy cells we need objetivity

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Dark Field – Pictures taken every 5 minutes EEVA Bright Field – Pictures taken every 10 minutes -GERI

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Three main things we can do with time lapse technology

Learn & Write a New Manual on Embryo Assessment

Support the Diagnosis ofBest Embryos to Transfer

Discover, Research & Develop

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414 Cryo‐transfers

435 KID Blastocysts

Initial ZP Thickness 

(µm)

Initial  area (µm2)

Initiation

expansion (min)

Minimum ZP Thickness 

(µm)

Maximumarea (µm2)

Morphokinetics of warmed blastocysts

Coello, Meseguer, Galan, Cobo ESHRE 2016

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Morphokinetics of warmed blastocysts

10

15

20

25

30

35

>9900 <9900Implantation rate (%)

Area (µm2)

Initial area

*

10

15

20

25

30

35

<18 um >18 um

Implantationrate

(%)

ZP thickness (µm)

Maximun ZP thickness

*

10

20

30

40

50

>19140 <19140Implantation rate (%)

Area (µm2)

Final Area

*

10

15

20

25

30

35

40

<12 um >12 um

Implantationrate

(%)

ZP thickness (µm)

Minimum ZP thickness

*

Coello, Meseguer, Galan, Cobo ESHRE 2016

PÁG.10Athayde Wirka et al. Fertil & Steril, 2014

Identify atypical phenotypes; Abnormal-cleavage

DC 1-3 Not DC 1-3

Implantationrate

2,9%

28,7%

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• AC1 and AC2 embryos are often selected for Day 3 transfer (28.6%)

• AC embryos are often good quality (46.9% 6-10 cells, ≤10% frag)

• Morphology is unable to detect AC embryos

• Implantation Rate: 3.7%

Abnormal Cleavage = Direct Cleavageconfirmation by automatic time-lapse

Blast Rate

ImplRate

Control (n=524) 43% 18%

With AC (n=115) 12% 4%

p-value <0.0001 0.05

Athayde Wirka et al. Fertil & Steril, In Press

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Identify atypical phenotypes; Blastocyst Contraction

Incidence; 19,1% of blastocysts contraction

*

J Marcos and M Meseguer 2015 Human Reporduction in Press

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Multinucleation

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1126 KID embryos analyzed

Nucleation status n %

NO MN 647 57.47

MN 479 42.53

Nucleation status n %

NO MN 963 85.6

MN 163 14,4

MN at 2 cells 127 77,9%

De novo MN 36 22,1%

reversibilityof =

multinucleation

(479-127)

479 = 73.4%

Multinucleation

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Multinucleation

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tEB

(107,94-112.99h)

tEB

(107,94-112.99h)

S3 R.O (0-3h)S3 R.O (0-3h)

A

72,2 %

A

72,2 %

B

66,2%

B

66,2%

S3 R.O (0-3h)S3 R.O (0-3h)

C

55,8%

C

55,8%

D

39,7%

D

39,7%

S3 = t8-t5

tEB

0

10

20

30

40

50

60

70

80

A B C D

72.266.2

55.8

39.7

Implantation rate

Tasas de implantación basadas en las diferentes categorías

Motato and Meseguer, ESHRE (2014)

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Blastocyst Selection Morphology vs Morphokinetics

tEB

(≤112.99h)

A

S3 R.O (≤ 3.0-5.67h)

A

72,2 %

B

66,2%

B

S3 R.O (≤ 3.0-5.67h)

C

55,8%

D

39,7%

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Blastocyst Selection Morphology vs Morphokinetics

Morphokinetics is superior than Morphology for blastocyst selection, when both combined morphology does not relate

with implantation

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Eeva Model; Bastocyst prediction

Eeva Morpho

HIgh High-Med Med-High Low

yes no

yes nono yes

cc2

9.33-11.47

s2

0-1.73h

s2

0-1.73h

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High

n= 158

• 46,8%*p<0,05

Medium

n= 114

• 36,8%

Low

n= 249

• 27,7%

Implantation rates according to Eeva categories: All transferred KID embryos n= 521

Implantation rates andEeva

B Aparicio-Ruiz and M Meseguer 2016 ESHRE

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Implantation rates according to Eeva categories: Logistic Regression analysis

Aparicio-Ruiz and Meseguer 2016 ESHRE

Model effect values OR p value

EEVA HIGH versus LOW 2.24 (1.44-3.47) >0.001

Day of Transfer Day 5 versus Day 3 2.17 (1.43-3.28) >0.001

Oocyte source Autologous versus Donation

3.17 (1.51-5,97) <0.001

Embryo morphology ASEBIR A vs. C 1.45 (0.79-2.65) ns

Eeva Algorithm is related with implantation potential independently of day of transfer, oocyte quality or

conventional embryo morphology which is not related with outcome when eeva is used

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Hypothesis

More Observations Better Selection

Less DisturbanceBetter Development

IndependentHEPA

particle filter

Accurate Temp Control (4 indep sensors)

Independent IncubatorsTrigasmixing

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Randomized Control Trial

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No NW data retrieved 1 4Time-lapse(N=216)*

Standard Incubator(N=162)

P

NewBorn delivery Rate 216/438 (49.3%) 162/405 (40.0%) 0.011Twins delivery 67/216 (31.0%) 40/162 (24.7%) NS

Female neonates 114 (45.2%) 51 (63.0%) <0.006

Birth weight (g) 3163 (3035-3292) 3074 (2913-3236) NS

Low birth weight (<2500g) 19 (12.8%) 15 (12.3%) NS

Very low birth weight (<1500g)

3 (2.0%) 3 (2.4%) NS

Neonatal height (cm) 50.3 (49.6-50.9) 49.7 (48.9-50.4) NS

Birth defects 1 (0.6%) 0 NS

F Insua and M Meseguer 2015 ESHRE

No comparable study on single point morphology observation

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=128)Mean CI95% Mean CI95% p-value[h] [h] [h] [h]

t4 38.8 37.9-39.6 40.5 39.6-41.4 0.009

t8 59.1 57.4-60.8 62.4 60.7-.64.2 0.008

tM 85.4 83.4-87.4 91.3 89.5-93.2 0.001

tSB 100.5 98.3-102.5 104.9 102.9-106.7 <0.001

tB 113.1 110.5-115.6 117.2 114.8-119.6<0.001

s2 1.14 0.8-1.5 2.15 1.6-2.70.001

Rienzi and M Meseguer 2016 Fertility (Submitted)

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tM

s2= t4-t3

Rienzi and M Meseguer 2016 Fertility (Submitted)

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Implantation in the categories of the hierarchical classification tree model applied in the

time lapse monitoring system, results are only referred to those euploid embryos with

known implantation (KID embryos). Xi Square p value=0.046

Blastocyst Category n Total n implanted % and CI95%

A 53 26 49.1 (35.7-61.6)

B 17 5 29.4 (7.7-51.1)

C 23 4 17.4 (1.9-32.9)

D 11 3 27.3 (1.0-53.6)

Distribution of euploid transferred embryos according to morphological categories

(ASEBIR). Xi Square p value=0.201

Blastocyst Category n Total n Implanted % and CI95%

A 55 23 41.8 (28.8-54.84)

B 40 14 35.0 (20.2-49.8)

C 9 1 11.1 (0.0-31.62)

D - - -

ROC CURVE VALUE AUC= 0.653

ROC CURVE VALUE AUC= 0.577

Rienzi and M Meseguer 2016 Fertility (Submitted)

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Implantation rates according to morphology and morphokinetics: Logistic Regression analysis

Model effect values OR p value

Morphokinetics A versus D 4.12 (1.39-12.24) >0.001

Maternal Age 1 year old 1.03 (1.43-3.28) ns

Embryo morphology ASEBIR A vs. C 1.19 (0.23-6.19) ns

Morphokinetics is superior than Morphology for blastocyst selection, when both combined morphology

does not relate with implantation

Rienzi and M Meseguer 2016 Fertility (Submitted)

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Fragmentation pattern

Bioanalytics on embryo development dynamics

Development of sub-cellular structures

Cellular shape dynamics

Computer vision can help us “see beyond”

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Computer vision can help us “see beyond”

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It is not easy to call attention of many embryologist like in this picture….

The evolution of TL incubators

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Water tank

• Six cameras with independentincubators

The evolution of TL incubators

LED- Orange led

Camera+Dish position

Second Dish Position Camera+Dish position

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One microscope per chamberOne microscope per chamber

11 focal planes of 96 micro-wells every 5 min11 focal planes of 96 micro-wells every 5 min

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Patient Name

The evolution of TL incubators

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4.9 MP Cameras (2560 x 1928)

4.9 MP Cameras (2560 x 1928)

Geri

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The evolution of Software analysis in TL

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The evolution of Software analysis in TL

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Question

If you have a time-lapse system available for clinicaluse in your lab, which type of patients/strategy will bebenefited/associated:

a) Patients with a previous IVF failure

b) Normal or high responder patients with more than 8embryos available to improve embryo selection

c) By using its blastocyst prediction abilities I will use it asa strategy to reach similar outcome than blastocysttransfer.

d) All patients would be benefitted due to the reducedembryo manipulation and improved culture conditions.

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Thanks for your attention

Marcos Meseguermarcos.meseguer@ivi.es