title: Extravillous Trophoblast Migration and Invasion Assay.
abstract: Extravillous trophoblast (EVT) migration and invasion through the decidualized endometrium is essential to successful placentation. SGHPL-4 cells, an EVT cell line derived from first trimester placenta, is a widely used model of cytotrophoblast differentiation into an invasive phenotype. Here we describe a quantitative cell migration assay that can be modified to also measure cell invasion. SGHPL-4 cells were seeded into BD Fluoroblok cell culture inserts constructed with an 8 μm porous membrane and allowed to migrate towards epidermal growth factor, a known chemoattractant for EVTs. To assess EVT invasion, Fluoroblok inserts were first coated with Matrigel, a basement membrane matrix. SGHPL-4 cells were labeled with calcein AM and cells that had invaded and/or migrated across the membrane were quantified by a bottom-reading fluorescence plate reader. The advantage of the Fluoroblok inserts over other migration/invasion assays is that they allow nondestructive detection of migrated cells.
authors.affiliation: Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, USA.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Angelova
authors.affiliation: Department of Biochemistry and Molecular Biology, Tulane University School of Medicine, New Orleans, USA.
authors.forename: Heather L
authors.initials: HL
authors.lastname: Machado
authors.affiliation: Department of Obstetrics and Gynecology, Tulane University School of Medicine, New Orleans, USA.
authors.forename: Kenneth F
authors.initials: KF
authors.lastname: Swan
authors.affiliation: Department of Obstetrics and Gynecology, Tulane University School of Medicine, New Orleans, USA.
authors.forename: Cindy
authors.initials: C
authors.lastname: Morris
authors.affiliation: Department of Obstetrics and Gynecology, Tulane University School of Medicine, New Orleans, USA.
authors.forename: Deborah E
authors.initials: DE
authors.lastname: Sullivan