Bone marrow concentrate for autologous transplantation in minipigs

M. Herten; J. P. Grassmann; M. Sager; L. Benga; J. C. Fischer; M. Jäger; M. Betsch; M. Wild; M. Hakimi; P. Jungbluth

doi:10.3415/VCOT-11-11-0165

Veterinary and Comparative Orthopaedics and Traumatology, Table of Contents

Vet Comp Orthop Traumatol 2013; 26(01): 34-41
DOI: 10.3415/VCOT-11-11-0165

Original Research

Schattauer GmbH

Bone marrow concentrate for autologous transplantation in minipigs

Characterization and osteogenic potential of mesenchymal stem cells

Authors

M. Herten

¹Department of Orthopaedics, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
J. P. Grassmann

²Department of Trauma and Hand Surgery, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
M. Sager

³Central Animal Research Facility, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
L. Benga

³Central Animal Research Facility, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
J. C. Fischer

⁴Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
M. Jäger

⁵University Duisburg-Essen, University Hospital, Department of Orthopaedics, Essen, Germany
M. Betsch

³Central Animal Research Facility, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
M. Wild

²Department of Trauma and Hand Surgery, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
M. Hakimi

²Department of Trauma and Hand Surgery, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany
P. Jungbluth

²Department of Trauma and Hand Surgery, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany

Abstract

Summary

Autologous bone marrow plays an increasing role in the treatment of bone, cartilage and tendon healing disorders. Cell-based therapies display promising results in the support of local regeneration, especially therapies using intra-operative one-step treatments with autologous progenitor cells. In the present study, bone marrow-derived cells were concentrated in a point-of-care device and investigated for their mesenchymal stem cell (MSC) characteristics and their osteogenic potential.

Bone marrow was harvested from the iliac crest of 16 minipigs. The mononucleated cells (MNC) were concentrated by gradient density centrifugation, cultivated, characterized by flow cytometry and stimulated into osteoblasts, adipocytes, and chondrocytes. Cell differentiation was investigated by histological and immunohistological staining of relevant lineage markers. The proliferation capacity was determined via colony forming units of fibroblast and of osteogenic alkaline-phosphatase-positive-cells.

The MNC could be enriched 3.5-fold in nucleated cell concentrate in comparison to bone marrow. Flow cytometry analysis revealed a positive signal for the MSC markers. Cells could be differentiated into the three lines confirming the MSC character. The cellular osteogenic potential correlated significantly with the percentage of newly formed bone in vivo in a porcine metaphyseal long-bone defect model.

This study demonstrates that bone marrow concentrate from minipigs display cells with MSC character and their osteogenic differentiation potential can be used for osseous defect repair in autologous transplantations.

Keywords

Mesenchymal stem cells - minipig - bone marrow concentrate - colony forming units

Full Text

References

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