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production), and Src inhibitors, PP2 and 10,12 conjugated linoleic
acid (CLA) rescue Ca2⁺ bursting in cells exposed to VEGF and
TNFa. We now investigate the effects of physiological (normal (NL)
vs PE) circulating levels of TNF, VEGF and IL-6 combined on Hu-
man umbilical vein endothelial cell (HUVEC) function and potential
rescue by CLA isomers 9, 11; 10,12, or a 50:50 mix.
STUDY DESIGN: HUVEC from NL pregnancies were grown to >90%
confluence and loaded with 10 uM Fura-2. Cells were stimulated
with 100 uM ATP alone, then washed prior to the NL pregnancy
cocktail (0.5 ng/ml VEGF, 0.1 ng/ml IL-6, and 0.03ng/ml TNFa) or
the PE cocktail (10ng/ml VEGF, 3.0 ng/ml IL-6, and 0.5 ng/ml
TNFa). After 30 min. cells were re-stimulation with ATP. Experi-
ments were then repeated with further pretreatment with 10 ng/mL
PP2 (control); 10,12 CLA alone; 9,11 CLA alone; or a 50:50 CLA mix
(doses below). Ca2⁺ sustained-phase burst numbers were compared
to internal control results.
RESULTS: While the NL cocktail had little effect on Ca2+ bursting, the
PE cocktail achieved 61% inhibition of function. Pretreatment with
PP2; 10, 12 CLA, and 10 uM CLA mix all showed significant recovery
(p<0.05) in sustained-phase Ca2⁺ bursting in HUVEC cells exposed to
the PE cocktail. Some lesser inhibition was noted in the NL pregnancy
cocktail group with the addition of PP2 and CLA 10 uM mix.
CONCLUSION: PE effects on endothelial cell function can be mimicked
by the combined addition of VEGF, TNFa, and IL-6 at levels found in
PE patients. 10, 12 CLA alone or in a 50:50 mix with 9,11 at 10 uM
improves these inhibitory effects in Ca2⁺ bursting. A 50:50 mix is
already FDA approved for use in pregnancy. Further studies of CLA
or another Src inhibitor, may lead to potential trials of prophylactic
and therapeutic treatments for PE pregnancies.
Rescue of PE Cocktail Inhibition
Agent Rescue
PP2 + the development of new drugs through the novel technology for
10,12 CLA 50 uM +
9,11 CLA 50 uM + Pregnancy-Associated Hypertension <33 weeks
50:50 CLA Mix 50 uM
10,12 CLA 10 uM + for the Eunice Kennedy Shriver National Institute of Child Health and
9,11 CLA 10 uM
50:50 CLA Mix 10 uM ++
961 Trophoblast differentiation of patient-specific
induced pluripotent stem cells from normal and
preeclampttic pregnancy
Young-Han Kim1, JoonHo Lee1, Yun ji Jung1, Geum Joon Cho2,
Han Sung Hwang3, Sung-Rae Cho4
1 clusion criteria for this analysis were singleton, vertex, non-anom-
Department of Obstetrics and Gynecology, Severance Hospital, Yonsei
Universtiy Medical College, Seoul, Korea, Republic of, 2Department of
Obstetrics and Gynecology, College of Medicine, Korea University, Seoul,
Korea, Republic of, 3Department of Obstetrics and Gynecology, KonKuk
University College of Medicine, Seoul, Korea, Republic of, 4Department and
Research Institute of Rehabilitation Medicine, Severance Hospital, Yonsei
Universtiy Medical College, Seoul, Korea, Republic of
Supplement to JANUARY 2017 American Journal of Obstetrics & Gynecology
Poster Session V
OBJECTIVE: Preeclampsia is thought to begin in inadequate tropho-
blast invasion and deficient remodeling of uterine spiral arteries.
Therefore, we generated patient-specific induced pluripotent stem
(iPS) cells from human amniotic epithelial (HAE) cells of women
with normal pregnancy and preeclampsia. Our aim was to induce
trophoblast differentiation from iPS cells and investigate candidate
genes associated with possible mechanism of preeclampsia.
STUDY DESIGN: From each HAE cells at 3rd or 4th passage, we
generated patient-specific iPS cells using Neon transfection and
Sendai virus treatment. Characteristics of iPS cells were identified by
conventional methods. In order to induce trophoblast, we per-
formed two methods: EB formation and none EB. EBs from iPS cells
were cultured for 7 days, and with BMP4(100ng/ml) on Matrigel-
coated dishes for 4-8 days. For feeder free method, iPS cells were
directly transferred on Matrigel-coated dishes, and cultured with
BMP4(100ng/ml). Characteristics of trophoblast were identified by
colony morphology, and ICC using CK7 as a trophoblast marker.
Patient-specific trophoblast from iPS cells were isolated and total
mRNA from each cells were prepared. We performed transcriptome
analysis to investigate the candidate genes associated with the
possible pathophysiology of preeclampsia.
RESULTS: These iPS cells were similar to human embryonic stem
(ES) cells in morphology, and expressed alkaline phosphatase.
Expression of surface markers and gene expressions like human ES
cells were confirmed. In this study, we induced differentiation into
trophoblast using these iPS cells, and obtained 59 candidate genes
associated with possible mechanism of preeclampsia in trophoblast
from iPS cells between 1 normal and 2 preeclampsia patients.
CONCLUSION: In further studies, these trophoblasts could be used as a
disease model of preeclampsia, and for investigating functional dif-
ferences among trophoblasts between normal and preeclampsia. Our
approach could eventually contribute (1) to enable a broad under-
standing of the pathogenesis of preeclampsia, (2) to enable further
study of other obstetric complications and (3) to obtain sources for
drug screening and toxicology in vitro.
962 Induction of labor vs. pre-labor cesarean in
gestation: success rate, duration, and maternal and
neonatal outcomes
Sarah Anderson
Human Development Maternal-Fetal Medicine Units Network, Bethesda, MD
OBJECTIVE: Some obstetricians opt for pre-labor cesarean delivery
(CD) over labor induction (IOL) in patients with Pregnancy-Asso-
ciated Hypertension (PAH) severe enough to require early preterm
delivery due to concerns about prolonged or failed IOL and
increasing morbidity. We compared maternal and perinatal out-
comes by IOL vs. pre-labor CD in women requiring delivery at <33
weeks for PAH. We also quantified the duration and success rate of
IOL.
STUDY DESIGN: Secondary analysis of an observational study that
enrolled 115,502 women in 25 US hospitals from 2008-2011. In-
alous live fetus, PAH and indicated delivery between 24 and <33
wks’. Exclusions were 2 prior CD and other indications for CD.
Components of the Maternal Composite (MC) and Neonatal
Composite (NC) outcomes are listed in Table 2. Logistic regression
and propensity scores were used for covariate adjustment (see Table
2 footnote). For duration of IOL, the start of IOL was the time of
S543