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Differentiation of mesenchymal stem cells in heparin-containing hydrogels via coculture with osteoblasts

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Abstract

The therapeutic potency of delivered mesenchymal stem cells (MSCs) in tissue engineering applications may be improved by priming cells toward a differentiated state via coculture with native, differentiated cells prior to implantation; however, there is a lack of understanding in what may be the most efficacious method. The objective of this study was to investigate the role of negatively-charged heparin in priming hydrogel-encapsulated MSCs toward the osteoblastic lineage during coculture with a monolayer of osteoblasts in the absence of dexamethasone. MSCs encapsulated with higher amounts of heparin and cocultured with osteoblasts exhibited an over 36-fold increase in alkaline phosphatase activity and 13-fold increase in calcium accumulation by day 21, compared to MSCs cocultured with MSCs at the same heparin content. Moreover, hydrogels with higher amounts of heparin and cocultured with osteoblasts exhibited enhanced mineralization on the edges, suggesting that heparin may be important in sequestering osteoblast-secreted soluble factors, particularly on the surfaces of hydrogels. The ability of heparin to selectively interact with soluble positively-charged proteins from the surroundings was confirmed through protein labeling and microscopy. These results suggest that heparin-containing hydrogels as part of a coculture system can be utilized as a versatile platform to study and enhance priming of MSCs toward various cell types for a wide variety of regenerative medicine-based therapies.

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Acknowledgement

The authors would like to thank Ryan C. Adams for his assistance in histology.

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Authors and Affiliations

  1. W.H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 313 Ferst Drive, Atlanta, GA, 30332, USA

    Song P. Seto & Johnna S. Temenoff

  2. School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, GA, 30332, USA

    Maria E. Casas

  3. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA, 30332, USA

    Johnna S. Temenoff

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  1. Song P. Seto
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  2. Maria E. Casas
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  3. Johnna S. Temenoff
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Correspondence to Johnna S. Temenoff.

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This work was supported by the Department of Education Graduate Assistance in Areas of National Need (GAANN) fellowship to S.P.S., NIH Biomaterial Training Grant (T32-EB006343) to M.E.C., National Science Foundation CAREER Award (CBET-0746209), and an Aircast Foundation grant to J.S.T. Human MSCs used in this study were provided by the Texas A&M Health Science Center College of Medicine Institute for Regenerative Medicine at Scott & White through a grant from NCRR of the NIH (P40RR017447).

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Seto, S.P., Casas, M.E. & Temenoff, J.S. Differentiation of mesenchymal stem cells in heparin-containing hydrogels via coculture with osteoblasts. Cell Tissue Res 347, 589–601 (2012). https://doi.org/10.1007/s00441-011-1265-8

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