The Stem Cell Niche

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Methylguanine DNA Methyltransferase-Mediated Drug Resistance-Based Selective Enrichment and Engraftment of Transplanted Stem Cells in Skeletal Muscle†‡

  1. Antonio S. J. Lee1,
  2. Prathibha Kahatapitiya1,
  3. Belinda Kramer1,
  4. Josephine E. Joya2,
  5. Jeff Hook1,
  6. Renjing Liu2,
  7. Galina Schevzov1,3,
  8. Ian E. Alexander4,
  9. Geoff McCowage5,
  10. Didier Montarras6,
  11. Peter W. Gunning1,3,7,
  12. Edna C. Hardeman2,8,§,*

Article first published online: 5 FEB 2009

DOI: 10.1002/stem.28

Issue

STEM CELLS

STEM CELLS

Volume 27, Issue 5, pages 1098–1108, May 2009

Additional Information

How to Cite

Lee, A. S. J., Kahatapitiya, P., Kramer, B., Joya, J. E., Hook, J., Liu, R., Schevzov, G., Alexander, I. E., McCowage, G., Montarras, D., Gunning, P. W. and Hardeman, E. C. (2009), Methylguanine DNA Methyltransferase-Mediated Drug Resistance-Based Selective Enrichment and Engraftment of Transplanted Stem Cells in Skeletal Muscle. STEM CELLS, 27: 1098–1108. doi: 10.1002/stem.28

Author Information

  1. 1

    Oncology Research Unit, The Children's Hospital at Westmead, Westmead, New South Wales 2145, Australia

  2. 2

    Muscle Development Unit, Children's Medical Research Institute, Westmead, New South Wales 2145, Australia

  3. 3

    Department of Pediatrics and Child Health, University of Sydney, New South Wales 2006, Australia

  4. 4

    Gene Therapy Research Unit, The Children's Hospital at Westmead, Westmead, New South Wales 2145, Australia

  5. 5

    Department of Oncology, The Children's Hospital at Westmead, Westmead, New South Wales 2145, Australia

  6. 6

    CNRS URA 2578, Department of Developmental Biology, Pasteur Institute, 75724 Paris Cedex 15, France

  7. 7

    Department of Pharmacology, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia

  8. 8

    Department of Anatomy, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia

  1. §

    Telephone: +61-29-385-2471; Fax: +61-29-385-8016.

*Department of Anatomy, School of Medical Sciences, University of New South Wales, Sydney, NSW 2052, Australia

  1. †

    Author contributions: A.S.J.L.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; P.K.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; B.K.: conception and design, provision of study material or patients; J.E.J.: conception and design, collection and/or assembly of data, data analysis and interpretation; J.H.: collection and/or assembly of data; R.L.: collection and/or assembly of data; G.S.: data analysis and interpretation; I.E.A.: provision of study material or patients, final approval of manuscript; G.M.: conception and design, provision of study material or patients; D.M.: provision of study material or patients, final approval of manuscript; P.W.G.: conception and design, financial support, provision of study material or patients, data analysis and interpretation, manuscript writing, final approval of manuscript; E.C.H.: conception and design, financial support, provision of study material or patients, data analysis and interpretation, manuscript writing, final approval of manuscript. A.S.J.L. and P.K. contributed equally to this article. The following laboratories have contributed equally to this work: Oncology Research Unit and Muscle Development Unit, School of Medical Sciences, University of New South Wales.

  2. ‡

    First published online in Stem CellsExpress February 5, 2009

  3. §

    Telephone: +61-29-385-2471; Fax: +61-29-385-8016.

Publication History

  1. Issue published online: 4 MAY 2009
  2. Article first published online: 5 FEB 2009
  3. Accepted manuscript online: 5 FEB 2009 12:00AM EST
  4. Manuscript Accepted: 22 JAN 2009
  5. Manuscript Received: 2 NOV 2008

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Keywords:

  • Adult stem cells;
  • CD34+;
  • Muscle stem cells;
  • Skeletal muscle;
  • Stem cell transplantation;
  • Tissue regeneration

Abstract

Cell replacement therapy using stem cell transplantation holds much promise in the field of regenerative medicine. In the area of hematopoietic stem cell transplantation, O6-methylguanine-DNA methyltransferase MGMT (P140K) gene-mediated drug resistance-based in vivo enrichment strategy of donor stem cells has been shown to achieve up to 75%–100% donor cell engraftment in the host's hematopoietic stem cell compartment following repeated rounds of selection. This strategy, however, has not been applied in any other organ system. We tested the feasibility of using this MGMT (P140K)-mediated enrichment strategy for cell transplantation in skeletal muscles of mice. We demonstrate that muscle cells expressing an MGMT (P140K) drug resistance gene can be protected and selectively enriched in response to alkylating chemotherapy both in vitro and in vivo. Upon transplantation of MGMT (P140K)-expressing male CD34+ve donor stem cells isolated from regenerating skeletal muscle into injured female muscle treated with alkylating chemotherapy, donor cells showed enhanced engraftment in the recipient muscle 7 days following transplantation as examined by quantitative-polymerase chain reaction using Y-chromosome specific primers. Fluorescent in situ hybridization analysis using a Y-chromosome paint probe revealed donor-derived de novo muscle fiber formation in the recipient muscle 14 days following transplantation, with approximately 12.5% of total nuclei within the regenerated recipient muscle being of donor origin. Following engraftment, the chemo-protected donor CD34+ve cells induced substantial endogenous regeneration of the chemo-ablated host muscle that is otherwise unable to self-regenerate. We conclude that the MGMT (P140K)-mediated enrichment strategy can be successfully implemented in muscle. Stem Cells 2009;27:1098–1108

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