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Published Online: 17 August 2020

Human Adipose-Derived Mesenchymal Stromal/Stem Cell Spheroids Possess High Adipogenic Capacity and Acquire an Adipose Tissue-like Extracellular Matrix Pattern

Publication: Tissue Engineering Part A
Volume 26, Issue Number 15-16

Abstract

Adipose-derived mesenchymal stromal/stem cells (ASCs) represent a commonly used cell source for adipose tissue engineering. In this context, ASCs have routinely been cultured in conventional 2D culture and applied as single cell suspension for seeding onto scaffold materials or direct injection. However, this approach is associated with the loss of their intrinsic 3D microenvironment and leads to impaired regenerative capacity of the cells. Thus, the application of ASCs as self-assembled 3D spheroids with cells residing in their own matrix is an attractive alternative. However, characterization of the structural features and differentiation capacity of the spheroids is necessary to effectively apply them as building blocks in adipose tissue engineering. In this study, we focus on extracellular matrix (ECM) development in ASC spheroids, as well as adipogenic differentiation in comparison to conventional 2D culture using different induction protocols. Reproducible assembly of ASCs into spheroids was achieved within 24 h using the liquid overlay technique. Undifferentiated spheroids displayed a stromal ECM pattern, with fibronectin, collagen V, and VI as the main components. In the course of adipogenesis, a dynamic shift in the ECM composition toward an adipogenic phenotype was observed, associated with enhanced expression of laminin, collagen I, IV, V, and VI, similar to native fat. Furthermore, adipogenic differentiation was enhanced in spheroids as compared with 2D cultured cells, with the spheroids needing a distinctly shorter adipogenic stimulus to sustain adipogenesis, which was demonstrated based on analysis of triglyceride content and adipogenic marker gene expression. In summary, culturing ASCs as spheroids can enhance their adipogenic capacity and generate adipose-like microtissues, which may be a promising cell delivery strategy for adipose tissue engineering approaches.

Impact statement

Adipose-derived mesenchymal stromal/stem cells (ASCs) as a widely used cell source for adipose tissue engineering have been shown to be limited in their regenerative capacity when applied as single cells. As an alternative approach, the delivery as spheroids, consisting of cells in a 3D context, may be favorable. However, insights into extracellular matrix (ECM) development and efficient adipogenic differentiation are required for their effective application. In this study, we show that differentiated ASC spheroids develop an ECM, resembling native adipose tissue. Furthermore, the ASC spheroids exhibited a superior differentiation capacity as compared with conventional 2D culture, and required only a short adipogenic induction stimulus. Our results identify ASC-derived spheroids as an attractive cell delivery method for adipose tissue engineering approaches.

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Information

Published In

cover image Tissue Engineering Part A
Tissue Engineering Part A
Volume 26Issue Number 15-16August 2020
Pages: 915 - 926
PubMed: 32070231

History

Published online: 17 August 2020
Published in print: August 2020
Published ahead of print: 9 April 2020
Published ahead of production: 19 February 2020
Accepted: 13 February 2020
Received: 7 August 2019

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Christiane Hoefner
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Christian Muhr
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Hannes Horder
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Miriam Wiesner
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Katharina Wittmann
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Daniel Lukaszyk
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Katrin Radeloff
Department of Otorhinolaryngology, University of Würzburg, Würzburg, Germany.
Marc Winnefeld
Beiersdorf AG, Hamburg, Germany.
Matthias Becker
Institute for Medical Radiation and Cell Research, University of Würzburg, Würzburg, Germany.
Torsten Blunk
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.
Petra Bauer-Kreisel [email protected]
Department of Trauma, Hand, Plastic and Reconstructive Surgery and University of Würzburg, Würzburg, Germany.

Notes

Address correspondence to: Petra Bauer-Kreisel, PhD, Department of Trauma, Hand, Plastic and Reconstructive Surgery, University of Wuerzburg, Oberduerrbacher Straße 6, Wuerzburg 97080, Germany [email protected]

Disclosure Statement

No competing financial interests exist.

Funding Information

This work was supported by the German Research Foundation (DFG), collaborative research center SFB TR225, project number 326998133 (sub-project C02). Furthermore, the authors wish to acknowledge the financial support by the Interdisciplinary Center for Clinical Research Wuerzburg, project number D-321, to K.R. and T.B.

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