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Interferon production in human placental trophoblast subpopulations
Placenta (1993), 14, AI-A84
5th MEETING OF THE EUROPEAN PLACENTA GROUP UNIVERSITY OF MANCHESTER, UK 8-11 September 1993 ABSTRACTS INTERFERON PRODUCTION IN HUMAN PLACENTAL TROPHOBLAST SUBPOPULATIONS. G. Aboagye-Mathiesen 1, F. D. T6thl, 2, P. M. Petersenl, V. Zacharl and P. Ebbesenl. 1Department of Virus and Cancer, The Danish Cancer Society, The Science Park, Gustav Wiedsvej 10, 8000 Aarhus C, Denmark. 2Institute of Microbiology, Medical University, H-4012 Debrecen, Hungary. Human placental trophoblast subpopulations were isolated from first and third trimester placentae and were infected with Sendal virus to produce interfetons (IFNs). The magnitude of the IFNs produced were different among the trophoblast populations. Highly proliferating first trimester extravillous trophoblast, characterized by being weakly HLA positive and high secretion of hCG, produced higher levels (five fold) of IFN than the first and third trimester villous cytotrophoblasts. Syncytiotrophoblast cultures established from mononuclear cytotrophoblast cultures produced twofold more IFNs than the mononuclear cytotrophoblast cultures when infected with the same virus. The level of IFN produced were dependent on the type of trophoblast, the proliferation rate, the gestational age and the stage of differentiation of the trophoblasts. The high level of IFN production in the first trimester trophoblast populations as compared to the term mononuclear trophoblast and syncytiotrophoblasts and the diverse biological effects of IFNs together may suggest a role of trophoblast IFNs in a complex series of events in human pregnancy.
TRANSFECT1ON OF HUMAN TROPHOBLAST CELLS IN CULTURE: A USEFUL TOOL TO STU,DY TRANSCRIPTIONAL REGULATION OF HORMONE GENES. E. Alsat*, P. Jacquemin, C. Oury, A. Belayew, D. Evain-Bfion , and J. A. Martial. *Physiopathologie du Dtveloppeme'nt, CNRS-ENS, 46, rue d'Ulm, 75230Paris, France, and Laboratoire de Biologic Moltculaire et de Gtnie G6nttique, Institut de Chimie, B6, Universit6 de LiSge, 4000-Sart Tilman, Belgium. In vitro, human cytotrophoblasts aggregate and fuse together to form syncytiotrophoblast. This morphological differentiation is associated with the induction of specific hormonal productions such as placental lactogen (hCS) and growth hormone (hGH-V). In this study, we have established a method for efficient lipofection of these cells which allowed us to study changes in the transient expression of hCS gene mutants during the course of syncytiotrophoblast formation. The plasmid used was pCSB-CAT where the placenta-specific enhancer of the hCS-B gene has been introduced into a vector (pBLCAT2) containing the thymidine kinase promoter linked to the CAT reporter gene. When cells grown for two days in vitro were transfccted in optimal lipofection conditions using Lipofcctin (Gibco-BRL) with pCSB-CAT, the transient CAT expression observed was 30fold higher relative to the pBLCAT2 control. This transcriptional activity increased progressively during cyto-syncytiotrophoblast formation and might be related to the wellknown increase in hCS production with the time in culture of these cells. Therefore, human trophoblast ceils in primary culture offer a useful and physiological model to study transcriptional regulation of placental hormone genes.
5th MEETING OF THE EUROPEAN PLACENTA GROUP UNIVERSITY OF MANCHESTER, UK 8-11 September 1993 ABSTRACTS INTERFERON PRODUCTION IN HUMAN PLACENTAL TROPHOBLAST SUBPOPULATIONS. G. Aboagye-Mathiesen 1, F. D. T6thl, 2, P. M. Petersenl, V. Zacharl and P. Ebbesenl. 1Department of Virus and Cancer, The Danish Cancer Society, The Science Park, Gustav Wiedsvej 10, 8000 Aarhus C, Denmark. 2Institute of Microbiology, Medical University, H-4012 Debrecen, Hungary. Human placental trophoblast subpopulations were isolated from first and third trimester placentae and were infected with Sendal virus to produce interfetons (IFNs). The magnitude of the IFNs produced were different among the trophoblast populations. Highly proliferating first trimester extravillous trophoblast, characterized by being weakly HLA positive and high secretion of hCG, produced higher levels (five fold) of IFN than the first and third trimester villous cytotrophoblasts. Syncytiotrophoblast cultures established from mononuclear cytotrophoblast cultures produced twofold more IFNs than the mononuclear cytotrophoblast cultures when infected with the same virus. The level of IFN produced were dependent on the type of trophoblast, the proliferation rate, the gestational age and the stage of differentiation of the trophoblasts. The high level of IFN production in the first trimester trophoblast populations as compared to the term mononuclear trophoblast and syncytiotrophoblasts and the diverse biological effects of IFNs together may suggest a role of trophoblast IFNs in a complex series of events in human pregnancy.
TRANSFECT1ON OF HUMAN TROPHOBLAST CELLS IN CULTURE: A USEFUL TOOL TO STU,DY TRANSCRIPTIONAL REGULATION OF HORMONE GENES. E. Alsat*, P. Jacquemin, C. Oury, A. Belayew, D. Evain-Bfion , and J. A. Martial. *Physiopathologie du Dtveloppeme'nt, CNRS-ENS, 46, rue d'Ulm, 75230Paris, France, and Laboratoire de Biologic Moltculaire et de Gtnie G6nttique, Institut de Chimie, B6, Universit6 de LiSge, 4000-Sart Tilman, Belgium. In vitro, human cytotrophoblasts aggregate and fuse together to form syncytiotrophoblast. This morphological differentiation is associated with the induction of specific hormonal productions such as placental lactogen (hCS) and growth hormone (hGH-V). In this study, we have established a method for efficient lipofection of these cells which allowed us to study changes in the transient expression of hCS gene mutants during the course of syncytiotrophoblast formation. The plasmid used was pCSB-CAT where the placenta-specific enhancer of the hCS-B gene has been introduced into a vector (pBLCAT2) containing the thymidine kinase promoter linked to the CAT reporter gene. When cells grown for two days in vitro were transfccted in optimal lipofection conditions using Lipofcctin (Gibco-BRL) with pCSB-CAT, the transient CAT expression observed was 30fold higher relative to the pBLCAT2 control. This transcriptional activity increased progressively during cyto-syncytiotrophoblast formation and might be related to the wellknown increase in hCS production with the time in culture of these cells. Therefore, human trophoblast ceils in primary culture offer a useful and physiological model to study transcriptional regulation of placental hormone genes.