scientific program • symposia - American Society for Reproductive ...

ART, UROLOGY AND PATIENT EDUCATION
Moderators: Tien Cheng “Arthur” Chang, E.L.D., Ph.D., and
Marius Meintjes, D.V.M, Ph.D.
V-1 11:18 AM
NON-INVASIVE ASSESSMENT OF EMBRYO VIABILITY USING
NOVEL AUTOMATED IMAGING TECHNOLOGY.
B. Behr1, S. L. Chavez1,2,4, K. E. Loewke1,4, R. A. Reijo
Pera1,2. 1Department of Obstetrics and Gynecology,
Stanford University School of Medicine, Stanford, CA;
2Institute for Stem Cell Biology and Regenerative Medicine,
Stanford University School of Medicine, Stanford, CA;
3Department of Mechanical Engineering, Stanford
University, Stanford, CA; 4Auxogyn, Inc., Menlo Park, CA.
OBJECTIVE: Recently, we demonstrated that the success or
failure of human embryos in reaching the blastocyst stage
can be predicted by the 4-cell stage of development
with >93% sensitivity and specificity using three cell cycle
imaging parameters observed prior to embryonic genome
activation. Analysis of gene expression profiles indicated
that embryos predicted to develop to the blastocyst stage
differ in gene expression patterns from those that arrest
prior to blastocyst formation, suggesting that these imaging
parameters can be used as a non-invasive indicator of
the underlying molecular programs. Here, we describe the
development of novel time-lapse imaging technology that
can be used in a conventional incubator with automatic
image data processing for the prediction of embryo viability.
DESIGN: In order for the time-lapse microscope to fit in a
standard incubator, only the critical components of the
microscope are used and designed for a short optical path.
To ensure embryo light safety, the microscope is comprised
of a low power LED, high sensitivity camera and darkfield
illumination, the latter of which also provides high contrast
between the cell membranes and cytoplasm for easy cell
tracking. Extraction of the cell cycle parameters is achieved
by automated software and a cell tracking algorithm,
whereby each image is compared to all predicted
simulations and assigned a likelihood based on similarity to
achieve the best fit model.
MATERIALS AND METHODS: With the development of
this novel automated imaging technology and the
recent identification of the three non-invasive cell cycle
parameters that accurately predict blastocyst formation by
the 4-cell stage, there is great potential to translate these
scientific findings into IVF laboratories and improve embryo
selection in combination with other criteria currently used in
clinical practice.
Disclosures: SC and KL are now employees of Auxogyn,
Inc., which licensed intellectual property resulting from this
research. BB and RRP own stock in Auxogyn.
__________________________________________________________
V-2 11:25 AM
A NOVEL MECHANISM IN THE DEVELOPMENT OF HUMAN
EMBRYOS WITH A SINGLE PRONUCLEUS (PN) AND TWO
UNEVEN PN IDENTIFIED BY TIME-LAPSE CINEMATOGRAPHY.
Y. Mio, K. Yumoto, K. Iwata, A. Imajyo, Y. Iba. Reproductive
Centre, Mio Fertility Clinic, Yonago, Tottori Prefecture, Japan.
OBJECTIVE: Although human embryos with a single
pronucleus (1PN) or uneven two pronuclei (≥10 μl of
VIDEO PROGRAM
Tuesday, October 18, 2011 11:15 am – 1:00 pm
ASRM Video Session I
Chapin Theatre
95
difference in diameter) are clinically observed on 16 to 18
hours after assisted reproductive technologies (IVF/ICSI),
the detailed mechanism in the development of these
embryos are still unknown. Recently, we determined a novel
mechanism occurring embryos with 1PN or uneven 2PN after
ICSI using time-lapse cinematography (TLC).
DESIGN: The TLC of donated ICSI embryos from 109 couples
was performed between April 2002 and December 2009.
Each ICSI oocyte was placed in 5μl of pre-equilibrated
fertilization medium within a TLC chamber on the stage of
an inverted microscope. Images were captured every 2
minutes (50 ms exposure time) for approximately 40 hours. Of
109 mature oocytes that underwent ICSI, 3 of both embryos
with 1PN or uneven 2PN were obtained from the TLC study.
MATERIALS AND METHODS: In 3 embryos with 1PN, an
unknown material was extruded next to 2nd polar body
(PB) 15 minutes after the extrusion of 2nd PB and continued
to exist in the perivitelline space until completion of the 1st
cleavage, whereas the male PN formed in the center of the
cytoplasm at 4.2 hours after ICSI. Although 3 embryos with
uneven 2PN also extruded an unknown material next to 2nd
PB, these embryos formed small PN-like substance (likely 3rd
PB) within the unknown material in the perivitelline space.
Afterward, the small PN-like substance incorporated into
ooplasm followed by migration to the male pronucleus and
abutment of both pronuclei.
This study firstly demonstrated the novel phenomena during
the miotic division in human embryos. Further TLC study will
give us more information about this enigmatic phenomena.
__________________________________________________________
V-3 11:32 AM
TIME-LAPSE IMAGING OF TRIPRONUCLEAR EMBRYOS:
MECHANISMS OF FORMATION AND ABNORMAL
DEVELOPMENT.
R. S. Weinerman1, M. E. Fino1, Y. Kramer1, K. C. Gunsalus2,
C. McCaffrey1, N. Noyes1. 1NYU Fertility Center, New York
University School of Medicine, New York, NY; 2Center for
Genomics and Systems Biology, New York University, New
York, NY..
OBJECTIVE: This video presents examples of abnormal
embryo development in tri-pronuclear embryos originating
from IVF±ICSI as viewed using time-lapse microscopy (TLM).
The mechanisms of tri-pronuclear embryo formation are
explained and resulting developmental phenotypes are
highlighted. Specific topics reviewed include mitotic spindle
formation, early embryo fragmentation, and early embryo
arrest.
DESIGN: Our research laboratory routinely employs TLM
as a means to study early embryogenesis. Our standard
clinical IVF practice is to assess all oocytes for evidence
of fertilization and to document pronuclear status 18 h
post-insemination by either conventional insemination or
ICSI. Those zygotes containing ≥3 pronuclei are deemed
unsuitable for uterine replacement and are sometimes
designated for continued culture and monitoring using TLM,
if the patient had signed IRB-approved informed consent
for research on this material. These abnormal zygotes are
placed in a stage-top incubator and viewed using highdefinition
microscopy. Images are captured every 240-420