Analysis of the Intrinsic Self-Organising Properties of Mesenchymal Stromal Cells in Three-Dimensional Co-Culture Models with Endothelial Cells.

Julia Marshall,Amanda Barnes,Paul Genever

doi:10.3390/bioengineering5040092

Julia Marshall, Amanda Barnes + Show 1 more

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https://doi.org/10.3390/bioengineering5040092

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Journal: Bioengineering	Publication Date: Oct 26, 2018
Citations: 7	License type: CC BY 4.0

Affiliation: University of York

Abstract

Mesenchymal stem/stromal cells (MSCs) are typically characterised by their ability to differentiate into skeletal (osteogenic, chondrogenic and adipogenic) lineages. MSCs also appear to have additional non-stem cell functions in coordinating tissue morphogenesis and organising vascular networks through interactions with endothelial cells (ECs). However, suitable experimental models to examine these apparently unique MSC properties are lacking. Following previous work, we have developed our 3D in vitro co-culture models to enable us to track cellular self-organisation events in heterotypic cell spheroids combining ECs, MSCs and their differentiated progeny. In these systems, MSCs, but not related fibroblastic cell types, promote the assembly of ECs into interconnected networks through intrinsic mechanisms, dependent on the relative abundance of MSC and EC numbers. Perturbation of endogenous platelet-derived growth factor (PDGF) signalling significantly increased EC network length, width and branching. When MSCs were pre-differentiated towards an osteogenic or chondrogenic lineage and co-cultured as mixed 3D spheroids, they segregated into polarised osseous and chondral regions. In the presence of ECs, the pre-differentiated MSCs redistributed to form a central mixed cell core with an outer osseous layer. Our findings demonstrate the intrinsic self-organising properties of MSCs, which may broaden their use in regenerative medicine and advance current approaches.

Highlights

Traditional two-dimensional (2D) cell culture techniques have provided valuable insights into fundamental aspects of cell function
These mesenchymal stem/stromal cells (MSCs):MSC (50:50) spheroids formed over the same timescale as the MSC-endothelial cells (ECs) spheroids, but in contrast, the red and green labelled cells appeared uniformly distributed at all time points and aggregates of red-labelled cells were not observed (Figure 1b, lower panel)
In this study we developed models to investigate the interactions between MSCs and ECs in 3D culture and in osteo/chondrogenic differentiation states

Summary

Introduction

Traditional two-dimensional (2D) cell culture techniques have provided valuable insights into fundamental aspects of cell function. Sophisticated 3D co-culture models can provide tractable systems to examine the mechanisms controlling the organisation and interaction of heterotypic cells, and may be developed for more effective forms of cell-based therapy. The use of mesenchymal stem/stromal cells (MSCs) in therapy has primarily been targeted towards regenerative applications, largely through their ability to differentiate into bone and cartilage tissues [1,2,3]. Long bone development through endochondral ossification and osteochondral tissue repair both involve a coordinated interplay between ECs and mesenchymal cells, to generate vascularised bone tissue and avascular cartilage [16]. It is important that we have appropriate tools to explore the relationship between MSCs and ECs to expand basic understanding of MSC biology and develop therapeutic applications, for example by engineering vascularised replacement tissues [17], and high-throughput screening platforms [18]

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