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Endoglandular trophoblasts disintegrate and replace uterine glandular epithelium in vitro and in situ
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
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