SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 178A

178A

Reproductive Sciences Vol. 25, Supplement 1, March 2018

T-206
Characterization of Human Term Amniotic Fluid Stem Cells. Cara
D Dolin†, Michael K Chan, Ross Basch, Bruce K Young*. New York
University Langone Health, New York, NY, United States.
INTRODUCTION: Stem cells, undifferentiated cells with the ability
to self-replicate without differentiation, have great therapeutic potential
for genetic and autoimmune disease, malignancies and even whole
organ replacement. Currently, stem cells sources include bone marrow,
peripheral blood, umbilical cord cells and blood, induced pluripotent cells
and embryonic tissue. Each of these sources has limitations. Amniotic
fluid stem cells (AFSC) collected in the first half of pregnancy have been
identified as a source of stem cells with possible therapeutic potential.
The objective was to characterize term AFSC, specifically (1) characterize
AFSC behavior compared to mid-trimester AFSC behavior; (2) determine
if term AFSC can survive multiple passages and cryostorage; and (3)
confirm the potential for differentiation.
METHODS: Amniotic fluid was collected from uncomplicated
pregnancies at time of scheduled, term, cesarean delivery. Fresh samples
were cultured and underwent cell expansion, and aliquots were frozen.
Cell viability and proliferation studies were performed. Flow cytometry
was used to identify surface markers associated with stem cells, including
SSEA-4, CD-90 and TRA-1-60. AFSC were differentiated toward
neural lineage and monoclonal antibodies for Nestin, B-tubulin III and
glial fibrillary acidic protein (GFAP) used to confirm differentiation by
fluorescent microscopy. AFSC were differentiated toward osteocyte
lineage and stained for presence of alkaline phosphatase-producing
cells, and stained with Alizarin Red solution, to confirm differentiation
to osteocyte lineage by bright-field and phase-contrast microscopy,
respectively. Differentiation of AFSC to chondrocytes was performed and
monoclonal antibodies for aggrecan were used to confirm differentiation.
RESULTS: Amniotic fluid was collected from 8 women with mean age
of 33.9 years (SD 2.7) and mean gestation age of 39.0 weeks (SD 0.9).
Volumes up to 100 ml were easily collected. All samples grew in culture
with mean cell viability of 80% (SD 7%), and cell counts of 1.3 x 10^5
to 1.5 x 10^6 per ml. Proliferation studies revealed a mean doubling time
of 27 hours. AFSC were able to tolerate 10 passages without significant
changes, and cryostorage for up to 18 months, without loss of viability.
All cells demonstrated surface markers associated with stem cells.
All samples demonstrated differentiation into neural, osteocyte and
chondrocyte lineages.
CONCLUSION: AFSC collected at term cesarean delivery demonstrate
characteristics similar to mid-trimester AFSC. Cells are able to be cultured
and withstand cryostorage. AFSC can be differentiated into multiple
cell lineages. AFSC may represent a readily available, abundant, and
noncontroversial source of stem cells for therapeutic potential, and
warrant further study.

T-207
Common Environmental Exposures Altering Zinc Levels Leading
to Poor Oocyte Quality and Fertilization Rates. Charalampos
Chatzicharalampous†, Roohi Jeelani*, Sarah Aldhaheri*, Sasha Mikhael†,
Robert Morris*, Husam Abu-Soud*. Wayne State University, Detroit,
MI, United States.
INTRODUCTION: Infertility affects one in six couples and the cause
may not be always apparent. Recent studies have suggested that common
toxins can impact reproductive health. These may include everyday
materials such as paint thinners, environmental pollutants such as
acrolein and styrene and heavy metal chelators such as Dimercapto-1propanesulfonic acid (DMPS). Zinc, a biologically essential transitional
metal, maintains oocyte integrity and supports embryo development.
Zinc is accumulated during oocyte growth and is thought to be stored in
lipoproteins in preparation for embryonic development. Even minimal
exposure to toxins is thought to result in changes to zinc homeostasis
leading to oocyte impairment. Given their ability to enter cells even at
low doses (2 μM) we sought to investigate the impact of these toxins on
1) oocyte quality, as judged by changes in microtubule morphology (MT)
and chromosomal alignment (CH) of metaphase II mice oocytes; and 2)
on zinc levels and subsequent embryo development.

Scientific Abstracts

METHODS: Metaphase II mouse oocytes (n=180) were exposed to
increasing concentrations of acrolein, styrene and DMPS (0-200 μM) for
45min to 2h (per protocol) after which all oocytes were fixed, stained and
scored based on the MT and CH as an indicator of the oocyte's capacity
to sustain exposure and then compared to untreated controls. Intracellular
zinc content was measured by utilizing Zinquin ethyl ester and using
confocal microscopy. Subsequently, outcomes of oocyte fertilization
rates after incubation in these compounds were determined by following
cleavage rate and development to the morula and blastocyst stages at 24,
48 and 72h post insemination.
RESULTS: A statistically significant difference in poor scores for MT and
CH was found between groups treated with increasing concentrations of
toxins as compared to controls (p< 0.05). A decrease in intracellular zinc
was noted as a function of increasing concentrations. Images of embryo
morphology at 24, 48 and 72h (2 cell / 8 cell / morula), and blastocyst
rates after exposure (0-200 μM) and IVF were assessed. At 24h post
insemination, the majority of oocytes exposed to all compounds appeared
granular and failed to fertilize compared to controls. The cleavage rate
and the rates of embryo development at 48h were significantly lower in
the different exposure groups (p<0.05). Most treated zygotes exhibited
fragmentation and a large perivitelline space. The arrested embryos were
highly fragmented and atretic with a dark granular appearance.
CONCLUSION: This work shows, for the first time, the negative impact
of these environmental factors on reproductive health. The common
feature among the different toxins is their capacity to mediate zinc
deficiency and enhancement of ROS that leads to oxidative damage in
oocytes and, ultimately, infertility.

T-208
Cryopreservation of Excess Embryos is Associated with Increased
Utilization of IVF Services among Women with Private Insurance for
IVF. Kelsey Anderson†,1 Darcy Broughton†,2 Kenan Omurtag*,2 Barton
Hamilton*,3 Emily Jungheim*.2 1Barnes Jewish Hospital/Washington
University in St. Louis, Saint Louis, MO, United States; 2Washington
University in St. Louis - Division of Reproductive Endocrinology and
Infertility, Department of Obstetrics and Gynecology, Saint Louis, MO,
United States; 3Washington University in St. Louis Olin Business School,
Saint Louis, MO, United States.
INTRODUCTION: It is well-established that insurance coverage for IVF
improves IVF utilization, and that women with IVF insurance coverage
are more likely to return for additional IVF cycles if they fail IVF. Factors
associated with IVF utilization among women with private IVF insurance
coverage have not been explored. Our goal was to identify predictors of
returning for IVF after a failed cycle within this population.
METHODS: Women with private IVF insurance coverage who initiated
IVF in our center between 2001 and 2010 were included and followed
through 2014. Women using donor oocytes or a gestational carrier were
excluded. Covariates included age, BMI, race, history of prior live birth,
number of oocytes retrieved, excess embryos frozen, average distance
from our clinic, and average family income based on zip code of residence.
The outcome of interest was whether or not women returned for an
additional cycle of IVF if they failed to conceive with their initial cycle.
Standard univariate statistics were used to identify associations between
covariates and the outcome. Significant covariates were further analyzed
using logistic regression. All analyses were performed in SPSS.
RESULTS: Age, race and BMI were not significant in univariate analyses
whereas income ($82,491 vs. $72,912, p=0.029), distance from the clinic
(43 miles versus 101 miles, p=0.029), total number of oocytes retrieved
(12 versus 10, p=0.006) and excess embryos frozen (RR 1.5, 95% CI 1.21.85) were associated with women coming back for an additional cycle
of IVF after a failed attempt. The only factor that remained significant
after regression analysis was having excess embryos frozen (OR 2.7,
95% CI 1.5-4.8).
CONCLUSION: Freezing excess embryos is associated with increased
probability of returning for IVF treatment after a previously failed IVF
cycle among women with private insurance for IVF. Many insurance
plans do not cover embryo cryopreservation or storage fees. For example,
the mandate for IVF insurance coverage in Illinois includes up to four



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

SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - Cover1
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SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - Cover3
SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - Cover4
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2020
https://www.nxtbook.com/nxtbooks/sage/psychologicalscience_demo
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2020
https://www.nxtbook.com/nxtbooks/sage/fai_202009
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_august2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2019
https://www.nxtbook.com/nxtbooks/sage/fai_201909
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_july2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2019
https://www.nxtbook.com/nxtbooks/sage/canadianpharmacistsjournal_05062019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2019
https://www.nxtbook.com/nxtbooks/sage/sri_supplement_201903
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2018
https://www.nxtbook.com/nxtbooks/sage/tec_20180810
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_julyaugust2018
https://www.nxtbook.com/nxtbooks/sage/fai_201807
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2018
https://www.nxtbook.com/nxtbooks/sage/sri_supplement_201803
https://www.nxtbook.com/nxtbooks/sage/slas_discovery_201712
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_november2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_september2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_julyaugust2017
https://www.nxtbook.com/nxtbooks/sage/fai_supplement_201709
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_may2017
https://www.nxtbook.com/nxtbooks/sage/fai_201706
https://www.nxtbook.com/nxtbooks/sage/fai_201607
https://www.nxtbookmedia.com