Extracellular matrix surface regulates self-assembly of three-dimensional placental trophoblast spheroids.

Michael K Wong,Sarah A Shawky,Aditya Aryasomayajula,P Ravi Selvaganapathy,Sandeep Raha,Madeline A Green,Tom Ewart,Nils Cordes

doi:10.1371/journal.pone.0199632

Michael K Wong, Sarah A Shawky + Show 6 more

Open Access

https://doi.org/10.1371/journal.pone.0199632

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Journal: PloS one	Publication Date: Jun 25, 2018
Citations: 29	License type: CC BY 4.0

Affiliation: McMaster University, McMaster University Medical Centre

Abstract

The incorporation of the extracellular matrix (ECM) is essential for generating in vitro models that truly represent the microarchitecture found in human tissues. However, the cell-cell and cell-ECM interactions in vitro remains poorly understood in placental trophoblast biology. We investigated the effects of varying the surface properties (surface thickness and stiffness) of two ECMs, collagen I and Matrigel, on placental trophoblast cell morphology, viability, proliferation, and expression of markers involved in differentiation/syncytial fusion. Most notably, thicker Matrigel surfaces were found to induce the self-assembly of trophoblast cells into 3D spheroids that exhibited thickness-dependent changes in viability, proliferation, syncytial fusion, and gene expression profiles compared to two-dimensional cultures. Changes in F-actin organization, cell spread morphologies, and integrin and matrix metalloproteinase gene expression profiles, further reveal that the response to surface thickness may be mediated in part through cellular stiffness-sensing mechanisms. Our derivation of self-assembling trophoblast spheroid cultures through regulation of ECM surface alone contributes to a deeper understanding of cell-ECM interactions, and may be important for the advancement of in vitro platforms for research or diagnostics.

Highlights

The human placenta is pivotal in the growth and survival of the fetus during pregnancy due to its involvement in maternal-fetal exchange, immune and barrier protection, and endocrine regulation [1, 2]
The current study demonstrates that the nature of the extracellular matrix (ECM) alone impacts the selfassembly behaviour of trophoblast cells, and the expression profiles of genes related to differentiation and cell-ECM interaction, and functionally alter syncytial fusion and hormone secretion
As the sophistication of in vitro research grows through the incorporation of ECM biomaterials, so does the necessity to better characterize the biological response of cells involved

Summary

Introduction

The human placenta is pivotal in the growth and survival of the fetus during pregnancy due to its involvement in maternal-fetal exchange, immune and barrier protection, and endocrine regulation [1, 2]. There has recently been great interest in emulating placental barrier function utilizing in vitro models comprised of monolayers of trophoblast cells or more complex assemblies of multiple cell types referred to as microphysiological systems [3,4,5]. Many of these in vitro platforms are developed in the absence of the non-cellular. The funders outlined in the “Sources of Funding” had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript They only provided financial support in form of author’s salaries (MKW, SAS, AA, MAG) and research materials.

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