title: Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium.
abstract: The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments.
authors.affiliation: Women's Reproductive Health Research Center, Vanderbilt University Medical Center, Nashville, TN, USA.|Department of Pathology, Immunology and Microbiology, Vanderbilt University Medical Center, Nashville, TN, USA.
authors.forename: Juan S
authors.initials: JS
authors.lastname: Gnecco
authors.affiliation: School of Electronic and Electrical Engineering, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, UK. V.Pensabene@leeds.ac.uk.|School of Medicine, Leeds Institute of Biomedical and Clinical Sciences, University of Leeds, Leeds, UK. V.Pensabene@leeds.ac.uk.
authors.forename: Virginia
authors.identifier: http://orcid.org/0000-0002-3352-8202
authors.initials: V
authors.lastname: Pensabene
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, CA, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Li
authors.affiliation: Women's Reproductive Health Research Center, Vanderbilt University Medical Center, Nashville, TN, USA.
authors.forename: Tianbing
authors.initials: T
authors.lastname: Ding
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, CA, USA.
authors.forename: Elliot E
authors.initials: EE
authors.lastname: Hui
authors.affiliation: Women's Reproductive Health Research Center, Vanderbilt University Medical Center, Nashville, TN, USA.
authors.forename: Kaylon L
authors.initials: KL
authors.lastname: Bruner-Tran
authors.affiliation: Women's Reproductive Health Research Center, Vanderbilt University Medical Center, Nashville, TN, USA.|Department of Pathology, Immunology and Microbiology, Vanderbilt University Medical Center, Nashville, TN, USA.|Veteran Affairs Tennessee Valley Healthcare System, Nashville, TN, USA.
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Osteen
mesh_terms: D018929:Cell Culture Techniques; D004717:Endometrium / Q000098:blood supply* / Q000166:cytology / Q000378:metabolism*; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008567:Membranes, Artificial; D008955:Models, Cardiovascular; D016062:Porosity; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D008567:Membranes, Artificial
keywords: Endometrium; Microfluidic; Organs-on-a-chip; Porous membrane; Stroma