SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 251A

Scientific Abstracts

As compared to the control group, OSI was higher in POI and IOF
(POI; 23.7±3.3, IOF; 20.7±3.6, Control; 16.5±2.1, P<0.001). However,
BAPs were not significantly different between the three groups (POI;
2029.0±186.4 μmol/l, IOF; 2116.2±240.2 μmol/l, Control; 2078.5±157.4
μmol/l ).
CONCLUSION: d-ROM and OSI increased in the POI and IOF. Our
results suggest that d-ROM and OSI may be a useful risk marker for POI.

F-212

F-213
M1 Macrophages Are Involved in Folliculogenesis and M2
Macrophages Play an Essential Role for Successful Implantation.
Yosuke Ono†, Osamu Yoshino*, Shigeru Saito*. University of Toyama,
Toyama, Japan.
INTRODUCTION: Macrophages (MΦs) are known to be involved in
folliculogenesis, ovulation, and implantation. It is, however, unknown
which type of MΦs, M1 or M2, play more essential roles for reproduction.
We evaluated the difference of the role of M1 or M2 MΦs using transgenic
mouse.
METHODS: We used two types of diphtheria-toxin receptor transgenic
(DTR) mice; CD11c-DTR mice can deplete CD11c+M1MΦ and dendritic
cells (DCs), and CD206-DTR mice can deplete CD206+ M2MΦs by
DT treatment. DT were injected to decrease CD11c+ or CD206+ cells in
superovulated mice with PMSG+hCG. Oocytes from the oviducts were
used for in vitro fertilization and embryo transfer. DT was injected just
before pre prior to implantation period in CD206-DTR female mice mated
with Balb/c male mice.

251A

RESULTS: In CD11c-DTR mice, folliculogenesis was completely
impaired with ovarian hemorrhage and the CD34-positive cells,
endothelial cells, were significantly reduced. In CD206-DTR mice,
folliculogenesis was normal and the number of ovulation, fertilization
rate, and implantation rate were similar to those in WT. In CD206-DTR
mice, implantation failure was observed on embryonic day 4.5 (E4.5),
although the serum levels of estradiol and progesterone were similar to
those in WT mice. Furthermore, decidualization at E3.5 was histologically
impaired in CD206-DTR mice. This impairment was due to proliferation
failure of endometrial stromal cells, lower staining of Ki 67. These findings
suggested that the depletion of M2MΦs resulted in implantation failure
by impairment of endometrial receptivity for embryos.
CONCLUSION: M1MΦs are involved in folliculogenesis, and M2MΦs
are essential for implantation.

F-214
DNA Methylation is Associated with Ploidy Status, Delayed
Blastulation, and Patient Age in Human Embryos. Xin Tao†,2,3 Yiping
Zhan,3 Katherine Scott,3 Richard T Scott, Jr,1,3 Emre Seli.4 1IVI-RMA New
Jersey, Basking Ridge, NJ, United States; 2Seton Hall University, South
Orange, NJ, United States; 3The Foundation for Embryonic Competence,
Basking Ridge, NJ, United States; 4Yale School of Medicine, New Haven,
CT, United States.
INTRODUCTION: DNA methylation (DNAme) is a fundamental
epigenetic control mechanism that occurs by the addition of methyl (CH3)
groups to cytosine residues and guides differentiation during embryonic
development. We hypothesized that DNAme may be associated with
viability of human embryos and compared genome-wide DNAme profile
in human blastocysts with different ploidy status, developmental stage,
and maternal age.
METHODS: Two trophoectoderm (TE) biopsies from each of the
previously diagnosed euploid (n=10) and aneuploid (n=10) embryos
were analyzed using Whole Genome Bisulfite Sequencing (WGBS).
Bisulfite conversion was performed using EZ DNA Methylation-Direct
Kit (Zymo). Unmethylated E. coli and methylated pUC19 DNA were
added as negative and positive controls, respectively, to monitor bisulfite
conversion efficiency. Methylome sequencing libraries were constructed
using TruSeq DNA Methylation Library Prep (Illumina) with 18 cycles of
amplification. Sequencing was performed on Illumina HiSeq 2500 with
paired-end 150 bp reads, and sequencing reads were aligned to human
genome reference using Bismark software. Duplicates were removed,
unconverted reads were filtered, and genome-wide cytosine methylation
at single base resolution was determined. Statistical analysis was carried
out using a linear model to assess the relationship of DNAme levels with
ploidy status, maternal age (range 29.5 to 41.1), and time of blastulation
(day 5 [n=8] vs. day 6 [n=12]).
RESULTS: The average CpG coverage achieved by WGBS in TE
samples was 30% of the sites predicted in the genome. Analysis revealed
that 20%-30% CpG sites in TE samples were methylated, consistent
with levels reported in studies using animal models. The unmethylated
E.coli and methylated pUC19 DNA controls showed 1<% and >98%
methylation rate, respectively. The two TE samples from the same
embryo showed significantly higher similarity of overall methylation
rate compared to the unrelated embryos (p<0.0001), which demonstrated
the reproducibility and feasibility of assessing DNAme from blastocyst
TE biopsies. Aneuploid embryos showed significantly higher DNAme
levels compared to euploid embryos (p<0.005), and increased patient age
was correlated with elevated DNAme levels in blastocysts (p<0.0001).
In addition, blastocysts cryopreserved on day 6 had significantly higher
DNAme compared to those that were cryopreserved on day 5 (p<0.0001).
CONCLUSION: DNA methylation levels detected in trophoectoderm
biopsies from human blastocysts correlate with ploidy status, maternal
age, and embryo growth characteristics, and may provide a foundation for
the development of epigenetic biomarkers of reproductive competence.

Friday Posters

"HCG Priming" for Diminished Ovarian Reserve Patients. Chaula
Mehta†*,2,3 Jigal Haas*,2,3 Rawad Bassil†,2,3 Eran Zilberberg†,2,3 Robert
Casper*,2,1,3 Ken Cadesky*,2,3 Dan Nayot*.2,3 1Samuel Lunenfeld Research
Institute, Toronto, ON, Canada; 2TRIO Fertility, Toronto, ON, Canada;
3
University of Toronto, Toronto, ON, Canada.
INTRODUCTION: Diminished ovarian reserve (DOR) is a major
contributing factor to IVF failure. In contrast, patients with PCOS have
a high ovarian reserve and respond well to ovarian stimulation. Priming
with androgen supplementation has shown some benefit in increasing
the ovarian reserve in DOR patients, but current routes of administration
have systemic side effects. The hyperandrogenic state noted in PCOS
patients is related to LH dominant micro-environment leading to thecal
androgen production. We hypothesized that 'HCG priming' would
achieve a temporary PCOS-like condition by acting specifically on the
LH receptors, in essence increasing local androgen production without
any systemic side effects. The aim of this study is to evaluate the influence
of 1-2 months treatment with daily low dose HCG on the ovarian reserve
of patients with DOR. Although HCG has been studied during ovarian
stimulation cycles, this is the first study to evaluate its role during priming
for 1-2 months duration.
METHODS: A retrospective case series using HCG priming in patients
with DOR to determine the change in their cycle day 3 (CD3) ovarian
reserve tests: antral follicle count (AFC) and FSH levels. Fourteen patients
with DOR were consented to participate and received HCG 200IU SC
daily for 1-2 months.
RESULTS: Fourteen patients underwent HCG priming treatment for 1-2
months (age 36.3±3.8 years). The results were analyzed using a paired 't'
test. The average AFC pre and post HCG treatment was 6.4 +/- 3.6 and
8.8 +/- 3.3 respectively (p = 0.04). Only 8 patients had their FSH values
recorded prior to and after HCG priming and noted to be 10 IU +/- 5.1
and 4.7 IU +/- 3.9 respectively (p < 0.001). No patients experienced any
adverse side effects from the HCG medications.
CONCLUSION: HCG priming appears to improve the ovarian reserve as
noted by an increased AFC and a decreased CD3 FSH level, while having
no side effects. Daily HCG mimics a PCOS-like ovarian environment by
creating an LH-dominant response leading to localized hyperandrogenic
state. This is preliminary data that serves as a proof of principal. A
prospective study is currently underway to better evaluate the impact
of HCG priming.
*Figure(s) will be available online.

Reproductive Sciences Vol. 25, Supplement 1, March 2018
Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018
