Short term exposure of BMP4 created novel trophoblast progenitor cells from human pluripotent stem cells

s / Placenta 36 (2015) A1eA14 A5 expression. These cells were cultured in the presence of human serum with or without complement inactivation and cell proliferations were evaluated by WST assay. Results: In immunohistochemistry, CD59 expression is higher on eEVT than on EVT. RT-PCR, immunocytochemistry, and flow cytometry showed that isolated EVT and Swan71 express CD59 intensely, whereas the expressions in BeWo, JAR and JEG are hardly detectable. In WST assay, there are no significant differences between two groups. JPA2015-15. SHORT TERM EXPOSURE OF BMP4 CREATED NOVEL TROPHOBLAST PROGENITOR CELLS FROM HUMAN PLURIPOTENT STEM CELLS Katsuyuki Adachi , Ying Yang , Toshihiko Ezashi , Andrei Alexenko , Megan Sheridan , D.J. Schust , L.C. Schulz , Mitsuyoshi Amita , R.M. Roberts , Kei Kawana , Yutaka Osuga , Tomoyuki Fujii . Department of OB/GYN School of Medicine University of Tokyo, Japan; Division of Animal Sciences, University of Missouri, Columbia, USA; Department of Obstetrics, Gynecology, and Women's Health, University of Missouri, Columbia, USA; Department of Biochemistry, University of Missouri, Columbia, USA Introduction: Preeclampsia is characterized by a lack of spiral artery remodeling in early placentation. Because trophoblast invasion occurs in early pregnancy when access to human placental tissue is limited, there is a need for model systems for the study of trophoblast differentiation. Objective:To establish unique stem cells that allow efficient trophoblast differentiation from human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) METHODS: hESC and iPSC were transiently exposed to BMP4 (10 ng/ml), A83-01 (1 mM) an activin A signaling inhibitor and PD173074 (0.1 mM) an FGF receptor inhibitor in culture medium lacking FGF2 for 24-36 h and then followed by trypsin dissociation and recovery of colonies capable of growing on a gelatin substratum in standard medium for human PSCs at low but not high FGF2 concentrations. RESULTS: Acquired self-renewing cell lines can be propagated clonally on gelatin and are morphologically distinct from human PSC progenitors but still meet standard in vitro criteria for pluripotency. The cells expressed weakly for CDX2 and strongly for NANOG and have a distinct transcriptome profile from the human PSCs fromwhich they were derived. The colonies spontaneously differentiated along multiple lineages, including trophoblast when cultured in nonconditioned medium lacking FGF2. They responded to PD173074 in the absence of both FGF2 and BMP4 by conversion to trophoblast, and especially syncytiotrophoblast, whereas an A83-01/PD173074 combination favored increased expression of HLA-G, a marker of extravillous trophoblast. They formwelldifferentiated teratomas in immune-compromised mice that secrete human chorionic gonadotropin (hCG) into the host mouse and include small areas of trophoblast-like cells. CONCLUSION: The cell we established that BMP4 can prime ESC and iPSC to a self-renewing alternative state permissive for trophoblast development. JPA2015-16. INDUCED PLURIPOTENT STEM CELLS (IPS CELL) DIFFERENTIATES TO TROPHOBLAST Junya Kojima , Hidenori Akutsu , Hirotaka Nishi , Naoaki Kuji , Keiichi Isaka . Department of Obstetrics and Gynecology, Tokyo Medical University, Japan; National Center for Child Health and Development, Japan Objective: The induced pluripotent stem cells (iPS cell) was derived from somatic cells. The iPS cell had pluripotency which could differentiate to almost all cells, and had self-renewal. iPS cell was different from embryonic stem cells, because we could make from various patient’s somatic cells. There were many reports trying to differentiate this iPS cell into various cells of the body like osteocyte, nerve cells, and hepatocyte cells. In this study, we used iPS cell, and induced differentiation of trophoblast that formed placenta. Methods: We used iPS cells derived from human endometrial cells, and made embryoid body (EB) on the suspension culture plate. We cultured EB for one week and adhered it on the dish coated with an extracellular matrix ( ). Then we add bone morphogenetic protein 4 (BMP4) 10ng/ml, 100ng/ml and cultured for one week. After that we collected cells, extracted RNA and made cDNA. The expression of undifferentiation marker (Oct3/4), trophoblast marker (CDX2, GCM1, syncytinI, syncytinII, KLF6, and PPAR-g) were measured by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Results: The expression of CDX2, GCM1, and syncytinI slightly increased in the BMP4 10ng/ml addition group compared with the control group. But there was no significant difference. In BMP4 100ng/ml addition group, all gene expression decreased compared with the control group. Conclusion: In this study, we used BMP4 and tried differentiation to trophoblast from iPS cells based on various reports, but could not differentiate effectively. Differentiation to trophoblast contributed to clinical elucidation and treatment of obstetrical disease, like preeclampsia, fetal growth restriction, and trophoblastic disease. We wanted to develop a more effective cultural method in future. JPA2015-17. PLACENTAMEGALY: FREQUENCY, CAUSES, AND COMPLICATIONS Arizawa Masayoshi. Tokyo Metropolitan Ohtsuka Hospital, Japan Introduction: The normal weight for a trimmed placenta is between 300 and 500 grams. If the weight is over 600 grams this is known as placentamegaly. This is usually associated with gestational diabetes (GDM), gigantism, virus infection, and placental hemangioma. However, we did not previously know the frequency of placentamegaly. This study looked at frequency, and complications with mother’s health and baby’s development. Method: I measured the weight of 1549 singleton placentas over a period of five years. I measured this by taking off the umbilical cord and membrane and measuring only the placental weight. Results: I found 63 placentamegaly cases out of 1549 high-risk cases sent to the department of pathology, which is 4.1% I studied the problems in three divisions; mothers, fetus and placenta A) Mothers’ problems From 63 cases there were 10 GDM, 4 unexamined pregnancies, and 1 mother’s anemia B) Babies’ problems From 63 cases we found 10 cases of 21 trisomy, and 4 cases of gigantism over 4000g C) Placenta problem From 63 cases we found 10 cases of chorangiosis, 9 cases of CAM, 7 cases of meconium stain, 2 cases of hemangioma, placental abnormal shape, and one case of abruptio placentae. Conclusion: Frommy study of 63 cases of placentamegaly (over 600 gram placental weight), I found that we should not only consider gigantism or GDM but also unexamined pregnancy, chromosomal abnormality, placental tumor, virus and bacterial infection, foetus hypoxia, and placental abnormal shapes. JPA2015-18. PREDICTION OF ADHERENT PLACENTA IN PATIENTS WITH PLACENTA PREVIA USING ULTRASONOGRAPHY AND MAGNETIC RESONANCE IMAGING Kenji Tanimura , Yui Yamasaki , Yoshiko Ueno , Tetsuo Maeda , Yasuhiko Ebina , Masashi Deguchi , Mayumi Morizane , Hideto Yamada . Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Japan; Department of Radiology, Kobe University Graduate School of Medicine, Japan Objective: Adherent placenta is a life-threatening condition in pregnancy, and is often complicated by placenta previa. The aim of this