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.Electronic supplementary materialThe online version of this article (doi:10.1007/s10439-017-1797-5) contains supplementary material, which is available to authorized users.

Amnion membrane organ‐on‐chip: an innovative approach to study cellular interactions

“…Recreating the whole organ dynamics using OOCs is an idea that has been around for many years (36); however, only very recently has their usefulness been explored in the field of reproductive science. The female reproductive tract (37), placenta (38), and endometrium-on-chip (39) have been developed and used to study multiple aspects of reproductive health, which have highlighted the importance of cell-cell and cell-blood interactions in vitro. Although development of a fetal membrane-on-chip has been postulated (22), no work has been reported yet.…”

Decidual stromal cell‐derived PGE2 regulates macrophage responses to microbial threat

“…The microfluidic organ-on-a-chip devices were assembled using our previously described protocol 41 .…”

Development of a novel human recellularized endometrium that responds to a 28-day hormone treatment†

“…The 28-day hormone regimen that mimics the human menstrual cycle is one that is not conventionally used in vitro to study hormone responses. Recently, a 28-day model of human perivascular stroma and endothelial cells was established using fixed concentrations of oestradiol, medroxyprogesterone acetate, and cAMP [52]. The concentration of hormones used in our 28-day protocol was based on clinical data of serum levels of hormones in women during the menstrual cycle [53,54].…”