title: A three-dimensional culture system recapitulates placental syncytiotrophoblast development and microbial resistance.
abstract: In eutherians, the placenta acts as a barrier and conduit at the maternal-fetal interface. Syncytiotrophoblasts, the multinucleated cells that cover the placental villous tree surfaces of the human placenta, are directly bathed in maternal blood and are formed by the fusion of progenitor cytotrophoblasts that underlie them. Despite their crucial role in fetal protection, many of the events that govern trophoblast fusion and protection from microbial infection are unknown. We describe a three-dimensional (3D)-based culture model using human JEG-3 trophoblast cells that develop syncytiotrophoblast phenotypes when cocultured with human microvascular endothelial cells. JEG-3 cells cultured in this system exhibit enhanced fusogenic activity and morphological and secretory activities strikingly similar to those of primary human syncytiotrophoblasts. RNASeq analyses extend the observed functional similarities to the transcriptome, where we observed significant overlap between syncytiotrophoblast-specific genes and 3D JEG-3 cultures. Furthermore, JEG-3 cells cultured in 3D are resistant to infection by viruses and Toxoplasma gondii, which mimics the high resistance of syncytiotrophoblasts to microbial infections in vivo. Given that this system is genetically manipulatable, it provides a new platform to dissect the mechanisms involved in syncytiotrophoblast development and microbial resistance.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Cameron A
authors.identifier: 0000-0001-6590-1668
authors.initials: CA
authors.lastname: McConkey
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Elizabeth
authors.identifier: 0000-0002-7455-7616
authors.initials: E
authors.lastname: Delorme-Axford
authors.affiliation: The Biodesign Institute, Center for Infectious Diseases and Vaccinology, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Nickerson
authors.affiliation: Division of Infectious Diseases, The Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Kwang Sik
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA 15219, USA.; Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Science, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Yoel
authors.initials: Y
authors.lastname: Sadovsky
authors.affiliation: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors.forename: Jon P
authors.initials: JP
authors.lastname: Boyle
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA 15219, USA.; Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Science, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Carolyn B
authors.initials: CB
authors.lastname: Coyne
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D010920:Placenta / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology; D011247:Pregnancy; D014158:Transcription, Genetic; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology*
keywords: 3-D cell culture; Placenta; RWV bioreactor; Toxoplasma gondii; trophoblast