Distribution and Viability of Fetal and Adult Human Bone Marrow Stromal Cells in a Biaxial Rotating Vessel Bioreactor after Seeding on Polymeric 3D Additive Manufactured Scaffolds.

Anne M Leferink,Lorenzo Moroni,Clemens A Van Blitterswijk,Yhee-Cheng Chng

doi:10.3389/fbioe.2015.00169

Abstract

One of the conventional approaches in tissue engineering is the use of scaffolds in combination with cells to obtain mechanically stable tissue constructs in vitro prior to implantation. Additive manufacturing by fused deposition modeling is a widely used technique to produce porous scaffolds with defined pore network, geometry, and therewith defined mechanical properties. Bone marrow-derived mesenchymal stromal cells (MSCs) are promising candidates for tissue engineering-based cell therapies due to their multipotent character. One of the hurdles to overcome when combining additive manufactured scaffolds with MSCs is the resulting heterogeneous cell distribution and limited cell proliferation capacity. In this study, we show that the use of a biaxial rotating bioreactor, after static culture of human fetal MSCs (hfMSCs) seeded on synthetic polymeric scaffolds, improved the homogeneity of cell and extracellular matrix distribution and increased the total cell number. Furthermore, we show that the relative mRNA expression levels of indicators for stemness and differentiation are not significantly changed upon this bioreactor culture, whereas static culture shows variations of several indicators for stemness and differentiation. The biaxial rotating bioreactor presented here offers a homogeneous distribution of hfMSCs, enabling studies on MSCs fate in additive manufactured scaffolds without inducing undesired differentiation.

Highlights

The field of tissue engineering aims at applying the fundamentals of cell biology and materials engineering to construct replacements for damaged, diseased, or lost tissue (Langer and Vacanti, 1993)
In the cross-sectional view of the scaffolds, it could be observed that the number of human fetal MSCs (hfMSCs) and their distribution seemed to be enhanced when the constructs were cultured in the biaxial bioreactor (Figure 2D) compared to the scaffolds with hfMSCs cultured statically in a well plate (Figure 2B)
The biaxial rotating vessel bioreactor used in this study has shown to improve hfMSCs distribution and proliferation in 3D additive manufactured poly(ethylene oxide terephthalate) (PEOT)/poly(butylene terephthalate) (PBT) scaffolds

Summary

Introduction

The field of tissue engineering aims at applying the fundamentals of cell biology and materials engineering to construct replacements for damaged, diseased, or lost tissue (Langer and Vacanti, 1993). Bone marrow stromal cells in 3D bioreactor cultures geometry, porosity, and tailored mechanical properties can be obtained by additive manufacturing (Moroni et al, 2005) These scaffolds provide the necessary support for cells to attach, proliferate, and differentiate, and define the overall shape of the tissue engineered transplant. Despite successes in regenerating tissues with additive manufactured scaffolds optionally combined with different bioreactors, the highly organized open structure of such scaffolds poses still challenges in homogenously distributing cells and controlling their proliferation and differentiation capacities. This is even more important when mesenchymal stromal cell (MSCs) are used, as their capacity to adhere and be homogeneously distributed in 3D scaffolds has shown to be more demanding (Griffon et al, 2011)

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Conclusion