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A Method to Isolate and Culture Expand Human Dental Pulp Stem Cells

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Mesenchymal Stem Cell Assays and Applications

Part of the book series: Methods in Molecular Biology ((MIMB,volume 698))

Abstract

Dentinal repair in teeth occurs through the activity of specialized cells known as odontoblasts that are thought to be maintained by a precursor population associated with the perivascular cells within dental pulp tissue. We have previously isolated candidate dental pulp stem cells (DPSC) from adult human third molars, with the ability to generate clonogenic cell clusters (CFU-F: colony-forming units-fibroblastic), a high proliferation rate, and multi-potential differentiation in vitro. When cultured DPSC are transplanted into immunocompromised mice, they generated a dentin-like structure lined with human odontoblast-like cells that surrounded a pulp-like interstitial tissue, composed of collagen and a vascular network. The present protocol describes a methodology to generate highly purified preparations of human DPSC. This process involves the enzymatic digestion of fresh samples of human dental pulp tissue followed by the isolation of DPSC using magnetic bead cell separation, based on their expression of mesenchymal stem cell associated markers.

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Acknowledgements

This work was supported by National Health and Medical Council of Australia Project Grant #453599 and #453497.

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Authors and Affiliations

  1. Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science/Hanson Institute/CSCR, University of Adelaide, Adelaide, SA, Australia

    Stan Gronthos

Authors
  1. Stan Gronthos
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  2. Agnieszka Arthur
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  3. P. Mark Bartold
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  4. Songtao Shi
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Corresponding author

Correspondence to Stan Gronthos .

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Editors and Affiliations

  1. , Primary and Stem Cell Systems, Life Technologies, Frederick, 21704, USA

    Mohan Vemuri

  2. Cellular Dynamics International, Inc., Science Drive 525, Madison, 53711, Wisconsin, USA

    Lucas G. Chase

  3. Life Technologies, Executive Way 7335, Frederick, 21704, Maryland, USA

    Mahendra S. Rao

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Gronthos, S., Arthur, A., Bartold, P.M., Shi, S. (2011). A Method to Isolate and Culture Expand Human Dental Pulp Stem Cells. In: Vemuri, M., Chase, L., Rao, M. (eds) Mesenchymal Stem Cell Assays and Applications. Methods in Molecular Biology, vol 698. Humana Press. https://doi.org/10.1007/978-1-60761-999-4_9

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  • DOI: https://doi.org/10.1007/978-1-60761-999-4_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-998-7

  • Online ISBN: 978-1-60761-999-4

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