title: Isolation and characterisation of a novel trophoblast side-population from first trimester placentae.
abstract: The placenta is responsible for all nutrient and gas exchange between mother and baby during pregnancy. The differentiation of specialised placental epithelial cells called trophoblasts is essential for placental function, but we understand little about how these populations arise. Mouse trophoblast stem cells have allowed us to understand many of the factors that regulate murine trophoblast lineage development, but the human placenta is anatomically very different from the mouse, and it is imperative to isolate a human trophoblast stem cell to understand human placental development. Here we have developed a novel methodology to isolate a Hoechst side-population of trophoblasts from early gestation placentae and compared their transcriptome to differentiated trophoblast populations (cytotrophoblasts and extravillous trophoblasts) using microarray technology. Side-population trophoblasts clustered as a transcriptomically distinct population but were more closely related to cytotrophoblasts than extravillous trophoblasts. Side-population trophoblasts up-regulated a number of genes characteristic of trophectoderm and murine trophoblast stem cells in comparison to cytotrophoblasts or extravillous trophoblasts and could be distinguished from both of these more mature populations by a unique set of 22 up-regulated genes, which were enriched for morphogenesis and organ development and the regulation of growth functions. Cells expressing two of these genes (LAMA2 and COL6A3) were distributed throughout the cytotrophoblast layer at the trophoblast/mesenchymal interface. Comparisons to previously published trophoblast progenitor populations suggest that the side-population trophoblasts isolated in this work are a novel human trophoblast population. Future work will determine whether these cells exhibit functional progenitor/stem cell attributes.
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia j.james@auckland.ac.nz.
authors.forename: J L
authors.initials: JL
authors.lastname: James
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: D G
authors.initials: DG
authors.lastname: Hurley
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: T K J B
authors.initials: TK
authors.lastname: Gamage
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: T
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: R
authors.initials: R
authors.lastname: Vather
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: P
authors.initials: P
authors.lastname: Pantham
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: P
authors.initials: P
authors.lastname: Murthi
authors.affiliation: Department of Obstetrics and GynaecologyFMHS, University of Auckland, 85 Park Road, Grafton, Auckland 1023, New ZealandSystems Biology LaboratoryMelbourne School of Engineering, University of Melbourne, Parkville, Melbourne, Victoria 3010, AustraliaAuckland District Health BoardGrafton, Auckland 1023, New ZealandDepartment of Perinatal MedicinePregnancy Research Centre, and University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Obstetrics and GynaecologyThe Royal Women's Hospital, 20 Flemington Road, Parkville, Melbourne, Victoria, Australia.
authors.forename: L W
authors.initials: LW
authors.lastname: Chamley
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002824:Chorionic Villi / Q000254:growth & development* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D010920:Placenta / Q000166:cytology* / Q000378:metabolism; D010929:Placentation / Q000502:physiology*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger