- Email: [email protected]
Vaccinium angustifolium var. laevifolium House (Lowbush blueberry) leaf extract increases trophoblast migration
Abstracts / Placenta 34 (2013) A1–A99
Conclusions: Based on our in silico analyses, literature data and first follow-up in vitro validations we were able to define potentially interesting candidate transporters implicated in PE and/or IUGR. Thorough analyses on protein and functional level will reveal whether these targets could be of diagnostic or therapeutical interest. http://dx.doi.org/10.1016/j.placenta.2013.06.280
P2.115. ESTROGEN RECEPTOR-MEDIATED INDUCTION OF ABCG2 TRANSPORTER BY MITOXANTRONE IN RAT PLACENTAL TROPHOBLAST CELLS
A95
system. The two electrodes voltage clamp system was used for functional analysis of GLUT9 (a and b isoforms) expressed in Xenopus laevis oocytres. Results: We could show that uric acid is transported across a BeWo cell monolayer with a rate of 530 pmol/min at the linear stage. We could successfully overexpress and purify these transporters. Further we could show that the tail currents of GLUT9a and GLUT9b show different electrical properties at positive membrane potentials. Conclusions: the fact of the slow Uric acid transport in the placenta could indicate that in vivo the fetus is protected from short-term fluctuations of maternal uric acid serum concentrations. Further investigation of the GLUT9 transporter isoforms on molecular levels showed different electrophysiological properties, indicating that the N-terminal difference of both isoforms is not a simple localization signal as suggested so far. We could also show that GLUT9-mediated uric acid transport is chloride dependent.
Masatoshi Tomi, Kenji Oda, Tomohiro Nishimura, Emi Nakashima http://dx.doi.org/10.1016/j.placenta.2013.06.282 Keio University Faculty of Pharmacy, Minato-ku, Tokyo, Japan Objectives: Placental syncytiotrophoblast expresses ABCG2/Breast Cancer Resistance Protein for suppressing fetal transfer of diverse compounds including mitoxantrone and pheophorbide-a. Molecular mechanisms underlying Abcg2 induction by its substrates may play an important role in regulating the fetal disposition of drugs. The aim of this study is to clarify the regulatory mechanism of Abcg2 expression by its substrate, mitoxantrone, in placental trophoblast TR-TBT 18d-1 cells. Methods: Quantitative real-time PCR and western blot were performed to measure gene expression levels in TR-TBT 18d-1 cells. Bisulphite pyrosequencing was used for measuring methylated CpG level in the promoter region. Results: The expression of Abcg2 in rat placental TR-TBT 18d-1 cells treated with mitoxantrone at more than 1 mM for 24 hrs was increased, compared with that in non-treated cells, while 10 mM pheophorbide-a had no effect. Methylated CpG level in the promoter region of the Abcg2 gene was low and was not altered by the mitoxantrone treatment. On the other hand, the mitoxantrone treatment markedly increased the expression of estrogen receptor (ER) a and progesterone receptor (PR) B. Fulvestrant, an ER antagonist, attenuated the mitoxantrone-induced increase of Abcg2 mRNA expression, while mifepristone, a PR antagonist, had little effect. 17bEstradiol, an ER ligand, positively regulated the mitoxantrone-induced increase of Abcg2 expression. DNA demethylation by 5-aza-2-deoxycytidine treatment increased ERa expression, but the mitoxantrone treatment failed to facilitate the demethylation of ERa promoter in TR-TBT 18d-1 cells. Conclusion: The expression of Abcg2 in TR-TBT 18d-1 cells was increased by mitoxantrone exposure via the induction of ERa. Reference: Oda et al., J. Pharm. Sci., in press. http://dx.doi.org/10.1016/j.placenta.2013.06.281
P2.116. PLACENTAL URIC ACID TRANSPORT SYSTEM Benjamin P. Lüscher 1, Benjamin Clémençon 2, Daniel V. Surbeck 3, Xiao Huang 2, 4, Camilla Marini 1, 4, Matthias A. Hediger 2, Christiane Albrecht 2, Marc Baumann 3 Department of Clinical Research, Bern, Switzerland; 2 Institute for Biochemistry and Molecular Medicine, Bern, Switzerland; 3 Department of Obstetrics and Gynecology, Bern, Switzerland; 4 Graduate School for Cellular and Biomedical Sciences, Bern, Switzerland 1
Objectives: Materno-fetal trans-placental transport is crucial for the fetal wellbeing. Elevated uric acid serum levels are often associated with pre-eclampsia. Uric acid acts as a pro-oxidant at high levels and can cause several cardiovascular diseases. We speculated that the placental uric acid transport system is altered in pre-eclampsia. The aim of this study was to investigate the transport capacity of BeWo choriocarcinoma cells and to evaluate the electrical properties of the glucose transporter GLUT9, which transports predominantly uric acid. Methods: We used the choriocarcinoma cell line as a placental cell model system to measure uric acid transport in a trans-epithelial transport
P2.117. PROTEOMIC ANALYSIS OF THE TROPHOBLASTIC BASAL MEMBRANE FRACTION FROM BEWO CELLS Soo-young Oh 1, Jae Ryoung Hwang 2, Jung-Joo An 1, Suk-Joo Choi 1, Jung-Sun Kim 3, Jong-Hwa Kim 1, Yoel Sadovsky 4, Cheong- Rae Roh 1 1 Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; 2 Samsung biomedical research institute, Samsung Medical Center, Seoul, Republic of Korea; 3 Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; 4 Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, USA
Background: The human placental syncytiotrophoblast is a polarized epithelium which forms the main barrier between mother and fetus. Within this microstructure, the fetal facing, basal membrane (BM) of the syncytiotrophoblasts exerts a key role in directing maternal-fetal transport. We used proteomics to characterize proteins in the trophoblast BM. Methods: We isolated BM fraction from BeWo cells by cationic colloidal silica (CCS) method. The isolation of BM fraction was validated by Western blot and immunofluorescence using the known markers for basal membrane (ad.enyl cyclase, plasma membrane calcium pump), apical membrane (placental alkaline phosphatase), and mitochondria (cytochrome C). Mass spectrometric analysis was performed to identify proteins from the BM. We also investigated the effect of hypoxia on the expression of the identified proteins by Western blot and the subcellular localization of these proteins by immunofluorescence microscopy. Results: We successfully isolated the basal membrane fraction of BeWo cells using CCS method, which we confirmed by Western blot and immunofluorescence microscopy. We found 13 proteins which were highly enriched in the BM fraction, including voltage-dependent anion channel 1 (VDAC1), ribophorin II (RPN2), heme oxygenase-1 (HO-1), and Rab14. We found that hypoxia increased the expression of these BM proteins and enhanced their membrane localization in BeWo cells. Conclusions: Hypoxia increases the expression of VDAC1, RPN2, HO-1, and Rab14 and their basal membrane localization in BeWo cells, suggesting that these proteins may have a role in placental-fetal transport in vivo. http://dx.doi.org/10.1016/j.placenta.2013.06.283
P2.118. VACCINIUM ANGUSTIFOLIUM VAR. LAEVIFOLIUM HOUSE (LOWBUSH BLUEBERRY) LEAF EXTRACT INCREASES TROPHOBLAST MIGRATION Christina Ly 1, 2, Julien Yockell-Lelièvre 2, Lana Saciragic 5, Ammar Saleem 3, Jonathan Ferrier 3, 4, John Thor Arnason 3, Andrée Gruslin 2, 5
A96
Abstracts / Placenta 34 (2013) A1–A99
1 Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada; 2 Ottawa Hospital Research Institute, Ottawa, ON, Canada; 3 Centre for Research in Biotechnology and Biopharmaceuticals, University of Ottawa, Ottawa, ON, Canada; 4 The New York Botanical Garden, Institute of Economic Botany, Bronx, NY, USA; 5 Department of Obstetrics and Gynecology, The Ottawa Hospital, Ottawa, ON, Canada
Objectives: Leaf extracts from many species of Vaccinium L., such as Vaccinium angustifolium Ait. (Lowbush blueberry), have been used for decades by indigenous populations of Canada in the context of diabetes. In other systems, Vaccinium has been shown to promote cell migration for wound healing. Many placenta-mediated diseases are associated with altered trophoblast migration. Since V. angustifolium has never been studied in the context of placental biology, we wanted to determine the effects of this extract on trophoblast biology. Methods: Dose-response studies were performed by treating HTR-8/ SVneo cells, a human extravillous trophoblast cell line, with V. angustifolium var. laevifolium House for 24 hours. Cell migration, proliferation, and viability were assessed using a Boyden chamber migration assay, BrdU incorporation assay, and trypan blue exclusion test, respectively. Western blot analyses of time-course studies were used to detect changes in ERK and AKT phosphorylation upon treatment. Results: Cells treated with 20 ng/mL of the extract displayed a significant increase in migratory ability compared to the untreated group (p<0.001). At this concentration, the extract had no effect on cell proliferation or viability. Changes in ERK and AKT phosphorylation detected at 0, 30, 60, 90, and 120 min after treatment were not different between control and treated groups. Conclusion: At 20 ng/mL, V. angustifolium leaf extract increases cell migration with no effect on proliferation or cell death. ERK and AKT phosphorylation are not affected with treatment. Fractionation studies are underway in order to identify the active components of the extract and results will be presented. http://dx.doi.org/10.1016/j.placenta.2013.06.284
P2.119. ENDOGLANDULAR TROPHOBLASTS DISINTEGRATE AND UTERINE GLANDULAR EPITHELIUM IN VITRO AND IN SITU
REPLACE
Gerit Moser, Gregor Weiss, Monika Sundl, Martin Gauster, Berthold Huppertz Medical University of Graz, Graz, Austria Objectives: Invasion of extravillous trophoblasts into maternal tissues serves attachment of the placenta to the uterus on the one hand and nutrition of the embryo on the other hand. Endoglandular trophoblasts have been described as invading extravillous trophoblasts nearby, attached to and in uterine glands. It is tempting to speculate on their role in enabling histiotrophic nutrition of the embryo prior to the establishment of the uteroplacental circulation. The aim of this study is to investigate the presence of endoglandular trophoblasts during early placental development. Methods: Pieces of decidua parietalis (6-10 weeks gestation) were confronted and cocultured directly with villous explants from the same pregnancy. Confrontations were harvested after 72h, cryosectioned and processed for immunohistochemical single and/or double staining. These sections were compared with invaded and non-invaded first trimester placentation sites in situ. Antibodies against cytokeratin 7, HLA-G and vWF were used. Results: Endoglandular trophoblast invasion occurs in vitro and in situ. They invade into uterine glands and replace the uterine epithelium. Hence, accumulated detached glandular epithelial cells can be repeatedly observed in the glandular lumen. Additionally, in areas of trophoblast invasion the glandular epithelium seems to be completely disintegrated compared to glandular epithelium in the non-invaded parts of the decidua. Conclusion: Extravillous trophoblasts invade more structures in the maternal decidua than was known until today, rather than specifically focus on uterine
spiral arteries. Paracrine factors may be responsible for the disintegration of uterine glands in the maternal decidua, in the areas of invasion. http://dx.doi.org/10.1016/j.placenta.2013.06.285
P2.120. POSTPARTUM WOMEN'S PERSPECTIVES ON THE DONATION OF THE PLACENTA FOR SCIENTIFIC RESEARCH AND THERAPEUTIC APPLICATIONS IN CAMPINAS, SAO PAULO, BRAZIL Rebecca Scott Yoshizawa 1, Maria José Duarte Osis 2, Simony Lira do Nascimento 3, José Guilherme Cecatti 3, Ana Carolina Godoy 3, Suelene Coelho 3 Queen's University, Kingston, Ontario, Canada; 2 CEMICAMP, State University of Campinas, Campinas, Sao Paulo, Brazil; 3 CAISM, State University of Campinas, Campinas, Sao Paulo, Brazil 1
Background: Little is known about public perspectives of scientific and therapeutic uses of the placenta. Anthropological studies describe spiritual and cultural valuations of the organ in different cultural contexts, but the prevalence and compatibility with scientific uses are not known. Gaps in knowledge potentiate ethical and clinical problems regarding collection practices and applications. To address these gaps, we studied postpartum women's perspectives on placental donation. Methods: Conducted at the State University of Campinas in the CAISM maternity hospital, the study consisted of a cross-sectional survey of 384 women and semi-structured interviews with 12 women in the immediate postpartum. The survey had 27 questions regarding demographics, opinions of the placenta and its scientific uses, and donation practices. Surveys were analysed quantitatively while interviews were analysed qualitatively using grounded coding; results were compared. Results: The average age of respondents was 27. 56% had more than one child. 45% were Caucasian; 38% were mixed-race. 74% identified with a Christian faith. 52% had high school education or higher. 13% regarded the placenta as spiritually important. 72% felt that knowing what happens to the placenta after birth was somewhat or very important. 78% supported the use of the placenta in scientific research and medicine. 59% reported that consent to collect the placenta is very or somewhat important. 75% wanted to be asked to donate their placenta. 78% preferred to have their doctor invite them; only 7% preferred the researcher. 60% preferred to be approached during prenatal appointments. Interviews suggested women appreciate being part of research and that receiving information about studies is important to them. Conclusions: The placenta is not waste to be collected without involvement of women. Women support scientific and therapeutic uses of placentas and want to be included in decision-making. Women also desire information about the placenta and placenta science. http://dx.doi.org/10.1016/j.placenta.2013.06.286
P2.121. ANNEXIN 5 INVOLVEMENT IN HUMAN TROPHOBLAST FUSION Severine Degrelle 1,3, Pascale Gerbaud 1, 2, Anthony Bouter 4, Jean Guibourdenche 1, 5, Alain Brisson 4, Daniele Evain-Brion 1, 3, Guillaume Pidoux 1, 2 1 INSERM, UMR-S 767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris F-75006, France; 2 Université Paris Descartes, Paris F-75006, France; 3 PremUP, Foundation, Faculté des Sciences Pharmaceutiques et Biologiques, Paris F-75006, France; 4 UMR CNRS 5248 CBMN - Université Bordeaux 1, Pessac F-33600, France; 5 AP-HP, Biologie Hormonale, CHU Cochin, Paris F75014, France
Conclusions: Based on our in silico analyses, literature data and first follow-up in vitro validations we were able to define potentially interesting candidate transporters implicated in PE and/or IUGR. Thorough analyses on protein and functional level will reveal whether these targets could be of diagnostic or therapeutical interest. http://dx.doi.org/10.1016/j.placenta.2013.06.280
P2.115. ESTROGEN RECEPTOR-MEDIATED INDUCTION OF ABCG2 TRANSPORTER BY MITOXANTRONE IN RAT PLACENTAL TROPHOBLAST CELLS
A95
system. The two electrodes voltage clamp system was used for functional analysis of GLUT9 (a and b isoforms) expressed in Xenopus laevis oocytres. Results: We could show that uric acid is transported across a BeWo cell monolayer with a rate of 530 pmol/min at the linear stage. We could successfully overexpress and purify these transporters. Further we could show that the tail currents of GLUT9a and GLUT9b show different electrical properties at positive membrane potentials. Conclusions: the fact of the slow Uric acid transport in the placenta could indicate that in vivo the fetus is protected from short-term fluctuations of maternal uric acid serum concentrations. Further investigation of the GLUT9 transporter isoforms on molecular levels showed different electrophysiological properties, indicating that the N-terminal difference of both isoforms is not a simple localization signal as suggested so far. We could also show that GLUT9-mediated uric acid transport is chloride dependent.
Masatoshi Tomi, Kenji Oda, Tomohiro Nishimura, Emi Nakashima http://dx.doi.org/10.1016/j.placenta.2013.06.282 Keio University Faculty of Pharmacy, Minato-ku, Tokyo, Japan Objectives: Placental syncytiotrophoblast expresses ABCG2/Breast Cancer Resistance Protein for suppressing fetal transfer of diverse compounds including mitoxantrone and pheophorbide-a. Molecular mechanisms underlying Abcg2 induction by its substrates may play an important role in regulating the fetal disposition of drugs. The aim of this study is to clarify the regulatory mechanism of Abcg2 expression by its substrate, mitoxantrone, in placental trophoblast TR-TBT 18d-1 cells. Methods: Quantitative real-time PCR and western blot were performed to measure gene expression levels in TR-TBT 18d-1 cells. Bisulphite pyrosequencing was used for measuring methylated CpG level in the promoter region. Results: The expression of Abcg2 in rat placental TR-TBT 18d-1 cells treated with mitoxantrone at more than 1 mM for 24 hrs was increased, compared with that in non-treated cells, while 10 mM pheophorbide-a had no effect. Methylated CpG level in the promoter region of the Abcg2 gene was low and was not altered by the mitoxantrone treatment. On the other hand, the mitoxantrone treatment markedly increased the expression of estrogen receptor (ER) a and progesterone receptor (PR) B. Fulvestrant, an ER antagonist, attenuated the mitoxantrone-induced increase of Abcg2 mRNA expression, while mifepristone, a PR antagonist, had little effect. 17bEstradiol, an ER ligand, positively regulated the mitoxantrone-induced increase of Abcg2 expression. DNA demethylation by 5-aza-2-deoxycytidine treatment increased ERa expression, but the mitoxantrone treatment failed to facilitate the demethylation of ERa promoter in TR-TBT 18d-1 cells. Conclusion: The expression of Abcg2 in TR-TBT 18d-1 cells was increased by mitoxantrone exposure via the induction of ERa. Reference: Oda et al., J. Pharm. Sci., in press. http://dx.doi.org/10.1016/j.placenta.2013.06.281
P2.116. PLACENTAL URIC ACID TRANSPORT SYSTEM Benjamin P. Lüscher 1, Benjamin Clémençon 2, Daniel V. Surbeck 3, Xiao Huang 2, 4, Camilla Marini 1, 4, Matthias A. Hediger 2, Christiane Albrecht 2, Marc Baumann 3 Department of Clinical Research, Bern, Switzerland; 2 Institute for Biochemistry and Molecular Medicine, Bern, Switzerland; 3 Department of Obstetrics and Gynecology, Bern, Switzerland; 4 Graduate School for Cellular and Biomedical Sciences, Bern, Switzerland 1
Objectives: Materno-fetal trans-placental transport is crucial for the fetal wellbeing. Elevated uric acid serum levels are often associated with pre-eclampsia. Uric acid acts as a pro-oxidant at high levels and can cause several cardiovascular diseases. We speculated that the placental uric acid transport system is altered in pre-eclampsia. The aim of this study was to investigate the transport capacity of BeWo choriocarcinoma cells and to evaluate the electrical properties of the glucose transporter GLUT9, which transports predominantly uric acid. Methods: We used the choriocarcinoma cell line as a placental cell model system to measure uric acid transport in a trans-epithelial transport
P2.117. PROTEOMIC ANALYSIS OF THE TROPHOBLASTIC BASAL MEMBRANE FRACTION FROM BEWO CELLS Soo-young Oh 1, Jae Ryoung Hwang 2, Jung-Joo An 1, Suk-Joo Choi 1, Jung-Sun Kim 3, Jong-Hwa Kim 1, Yoel Sadovsky 4, Cheong- Rae Roh 1 1 Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; 2 Samsung biomedical research institute, Samsung Medical Center, Seoul, Republic of Korea; 3 Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; 4 Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, USA
Background: The human placental syncytiotrophoblast is a polarized epithelium which forms the main barrier between mother and fetus. Within this microstructure, the fetal facing, basal membrane (BM) of the syncytiotrophoblasts exerts a key role in directing maternal-fetal transport. We used proteomics to characterize proteins in the trophoblast BM. Methods: We isolated BM fraction from BeWo cells by cationic colloidal silica (CCS) method. The isolation of BM fraction was validated by Western blot and immunofluorescence using the known markers for basal membrane (ad.enyl cyclase, plasma membrane calcium pump), apical membrane (placental alkaline phosphatase), and mitochondria (cytochrome C). Mass spectrometric analysis was performed to identify proteins from the BM. We also investigated the effect of hypoxia on the expression of the identified proteins by Western blot and the subcellular localization of these proteins by immunofluorescence microscopy. Results: We successfully isolated the basal membrane fraction of BeWo cells using CCS method, which we confirmed by Western blot and immunofluorescence microscopy. We found 13 proteins which were highly enriched in the BM fraction, including voltage-dependent anion channel 1 (VDAC1), ribophorin II (RPN2), heme oxygenase-1 (HO-1), and Rab14. We found that hypoxia increased the expression of these BM proteins and enhanced their membrane localization in BeWo cells. Conclusions: Hypoxia increases the expression of VDAC1, RPN2, HO-1, and Rab14 and their basal membrane localization in BeWo cells, suggesting that these proteins may have a role in placental-fetal transport in vivo. http://dx.doi.org/10.1016/j.placenta.2013.06.283
P2.118. VACCINIUM ANGUSTIFOLIUM VAR. LAEVIFOLIUM HOUSE (LOWBUSH BLUEBERRY) LEAF EXTRACT INCREASES TROPHOBLAST MIGRATION Christina Ly 1, 2, Julien Yockell-Lelièvre 2, Lana Saciragic 5, Ammar Saleem 3, Jonathan Ferrier 3, 4, John Thor Arnason 3, Andrée Gruslin 2, 5
A96
Abstracts / Placenta 34 (2013) A1–A99
1 Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada; 2 Ottawa Hospital Research Institute, Ottawa, ON, Canada; 3 Centre for Research in Biotechnology and Biopharmaceuticals, University of Ottawa, Ottawa, ON, Canada; 4 The New York Botanical Garden, Institute of Economic Botany, Bronx, NY, USA; 5 Department of Obstetrics and Gynecology, The Ottawa Hospital, Ottawa, ON, Canada
Objectives: Leaf extracts from many species of Vaccinium L., such as Vaccinium angustifolium Ait. (Lowbush blueberry), have been used for decades by indigenous populations of Canada in the context of diabetes. In other systems, Vaccinium has been shown to promote cell migration for wound healing. Many placenta-mediated diseases are associated with altered trophoblast migration. Since V. angustifolium has never been studied in the context of placental biology, we wanted to determine the effects of this extract on trophoblast biology. Methods: Dose-response studies were performed by treating HTR-8/ SVneo cells, a human extravillous trophoblast cell line, with V. angustifolium var. laevifolium House for 24 hours. Cell migration, proliferation, and viability were assessed using a Boyden chamber migration assay, BrdU incorporation assay, and trypan blue exclusion test, respectively. Western blot analyses of time-course studies were used to detect changes in ERK and AKT phosphorylation upon treatment. Results: Cells treated with 20 ng/mL of the extract displayed a significant increase in migratory ability compared to the untreated group (p<0.001). At this concentration, the extract had no effect on cell proliferation or viability. Changes in ERK and AKT phosphorylation detected at 0, 30, 60, 90, and 120 min after treatment were not different between control and treated groups. Conclusion: At 20 ng/mL, V. angustifolium leaf extract increases cell migration with no effect on proliferation or cell death. ERK and AKT phosphorylation are not affected with treatment. Fractionation studies are underway in order to identify the active components of the extract and results will be presented. http://dx.doi.org/10.1016/j.placenta.2013.06.284
P2.119. ENDOGLANDULAR TROPHOBLASTS DISINTEGRATE AND UTERINE GLANDULAR EPITHELIUM IN VITRO AND IN SITU
REPLACE
Gerit Moser, Gregor Weiss, Monika Sundl, Martin Gauster, Berthold Huppertz Medical University of Graz, Graz, Austria Objectives: Invasion of extravillous trophoblasts into maternal tissues serves attachment of the placenta to the uterus on the one hand and nutrition of the embryo on the other hand. Endoglandular trophoblasts have been described as invading extravillous trophoblasts nearby, attached to and in uterine glands. It is tempting to speculate on their role in enabling histiotrophic nutrition of the embryo prior to the establishment of the uteroplacental circulation. The aim of this study is to investigate the presence of endoglandular trophoblasts during early placental development. Methods: Pieces of decidua parietalis (6-10 weeks gestation) were confronted and cocultured directly with villous explants from the same pregnancy. Confrontations were harvested after 72h, cryosectioned and processed for immunohistochemical single and/or double staining. These sections were compared with invaded and non-invaded first trimester placentation sites in situ. Antibodies against cytokeratin 7, HLA-G and vWF were used. Results: Endoglandular trophoblast invasion occurs in vitro and in situ. They invade into uterine glands and replace the uterine epithelium. Hence, accumulated detached glandular epithelial cells can be repeatedly observed in the glandular lumen. Additionally, in areas of trophoblast invasion the glandular epithelium seems to be completely disintegrated compared to glandular epithelium in the non-invaded parts of the decidua. Conclusion: Extravillous trophoblasts invade more structures in the maternal decidua than was known until today, rather than specifically focus on uterine
spiral arteries. Paracrine factors may be responsible for the disintegration of uterine glands in the maternal decidua, in the areas of invasion. http://dx.doi.org/10.1016/j.placenta.2013.06.285
P2.120. POSTPARTUM WOMEN'S PERSPECTIVES ON THE DONATION OF THE PLACENTA FOR SCIENTIFIC RESEARCH AND THERAPEUTIC APPLICATIONS IN CAMPINAS, SAO PAULO, BRAZIL Rebecca Scott Yoshizawa 1, Maria José Duarte Osis 2, Simony Lira do Nascimento 3, José Guilherme Cecatti 3, Ana Carolina Godoy 3, Suelene Coelho 3 Queen's University, Kingston, Ontario, Canada; 2 CEMICAMP, State University of Campinas, Campinas, Sao Paulo, Brazil; 3 CAISM, State University of Campinas, Campinas, Sao Paulo, Brazil 1
Background: Little is known about public perspectives of scientific and therapeutic uses of the placenta. Anthropological studies describe spiritual and cultural valuations of the organ in different cultural contexts, but the prevalence and compatibility with scientific uses are not known. Gaps in knowledge potentiate ethical and clinical problems regarding collection practices and applications. To address these gaps, we studied postpartum women's perspectives on placental donation. Methods: Conducted at the State University of Campinas in the CAISM maternity hospital, the study consisted of a cross-sectional survey of 384 women and semi-structured interviews with 12 women in the immediate postpartum. The survey had 27 questions regarding demographics, opinions of the placenta and its scientific uses, and donation practices. Surveys were analysed quantitatively while interviews were analysed qualitatively using grounded coding; results were compared. Results: The average age of respondents was 27. 56% had more than one child. 45% were Caucasian; 38% were mixed-race. 74% identified with a Christian faith. 52% had high school education or higher. 13% regarded the placenta as spiritually important. 72% felt that knowing what happens to the placenta after birth was somewhat or very important. 78% supported the use of the placenta in scientific research and medicine. 59% reported that consent to collect the placenta is very or somewhat important. 75% wanted to be asked to donate their placenta. 78% preferred to have their doctor invite them; only 7% preferred the researcher. 60% preferred to be approached during prenatal appointments. Interviews suggested women appreciate being part of research and that receiving information about studies is important to them. Conclusions: The placenta is not waste to be collected without involvement of women. Women support scientific and therapeutic uses of placentas and want to be included in decision-making. Women also desire information about the placenta and placenta science. http://dx.doi.org/10.1016/j.placenta.2013.06.286
P2.121. ANNEXIN 5 INVOLVEMENT IN HUMAN TROPHOBLAST FUSION Severine Degrelle 1,3, Pascale Gerbaud 1, 2, Anthony Bouter 4, Jean Guibourdenche 1, 5, Alain Brisson 4, Daniele Evain-Brion 1, 3, Guillaume Pidoux 1, 2 1 INSERM, UMR-S 767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris F-75006, France; 2 Université Paris Descartes, Paris F-75006, France; 3 PremUP, Foundation, Faculté des Sciences Pharmaceutiques et Biologiques, Paris F-75006, France; 4 UMR CNRS 5248 CBMN - Université Bordeaux 1, Pessac F-33600, France; 5 AP-HP, Biologie Hormonale, CHU Cochin, Paris F75014, France