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Cellular and Molecular Bioengineering

, Volume 11, Issue 2, pp 99–115 | Cite as

Generating Chondromimetic Mesenchymal Stem Cell Spheroids by Regulating Media Composition and Surface Coating

  • BanuPriya Sridharan
  • Amy D. Laflin
  • Michael S. Detamore
Article

Abstract

Introduction

Spheroids of mesenchymal stem cells (MSCs) in cartilage tissue engineering have been shown to enhance regenerative potential owing to their 3D structure. In this study, we explored the possibility of priming spheroids under different media to replace the use of inductive surface coatings for chondrogenic differentiation.

Methods

Rat bone marrow-derived MSCs were organized into cell spheroids by the hanging drop technique and subsequently cultured on hyaluronic acid (HA) coated or non-coated well plates under different cell media conditions. Endpoint analysis included cell viability, DNA and Glycosaminoglycan (GAG) and collagen content, gene expression and immunohistochemistry.

Results

For chondrogenic applications, MSC spheroids derived on non-coated surfaces outperformed the spheroids derived from HA-coated surfaces in matrix synthesis and collagen II gene expression. Spheroids on non-coated surfaces gave rise to the highest collagen and GAG when primed with medium containing insulin-like growth factor (IGF) for 1 week during spheroid formation. Spheroids that were grown in chondroinductive raw material-inclusive media such as aggrecan or chondroitin sulfate exhibited the highest Collagen II gene expression in the non-coated surface at 1 week.

Conclusion

Media priming by growth factors and raw materials might be a more predictive influencer of chondrogenesis compared to inductive-surfaces. Such tailored bioactivity of the stem cell spheroids in the stage of the spheroid formation may give rise to a platform technology that may eventually produce spheroids capable of chondrogenesis achieved by mere media manipulation, skipping the need for additional culture on a modified surface, that paves the way for cost-effective technologies.

Keywords

Mesenchymal stem cell Spheroids Growth factor Chondrogenesis Priming 

Notes

Compliance with Ethical Standards

The National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health under Award Number R01 AR056347 supported research reported in this publication. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We would also like to recognize support from the Kansas Bioscience Authority Rising Star Award. BanuPriya Sridharan, Amy D. Laflin, and Michael S. Detamore, have no financial or intellectual property conflicts to disclose. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

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Copyright information

© Biomedical Engineering Society 2017

Authors and Affiliations

  • BanuPriya Sridharan
    • 1
  • Amy D. Laflin
    • 2
  • Michael S. Detamore
    • 3
  1. 1.Bioengineering ProgramUniversity of KansasLawrenceUSA
  2. 2.Department of Chemical and Petroleum EngineeringUniversity of KansasLawrenceUSA
  3. 3.Stephenson School of Biomedical EngineeringUniversity of OklahomaNormanUSA

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