Fate of bone marrow mesenchymal stem cells following the allogeneic transplantation of cartilaginous aggregates into osteochondral defects of rabbits

The purpose of this study was to track mesenchymal stem cells (MSCs) labelled with internalizing quantum dots (i‐QDs) in the reparative tissues, following the allogeneic transplantation of three‐dimensional (3D) cartilaginous aggregates into the osteochondral defects of rabbits. QDs were conjugated with a unique internalizing antibody against a heat shock protein‐70 (hsp70) family stress chaperone, mortalin, which is upregulated and expressed on the surface of dividing cells. The i‐QDs were added to the culture medium for 24 h. Scaffold‐free cartilaginous aggregates formed from i‐QD‐labelled MSCs (i‐MSCs), using a 3D culture system with chondrogenic supplements for 1 week, were transplanted into osteochondral defects of rabbits. At 4, 8 and 26 weeks after the transplantation, the reparative tissues were evaluated macroscopically, histologically and fluoroscopically. At as early as 4 weeks, the defects were covered with a white tissue resembling articular cartilage. In histological appearance, the reparative tissues resembled hyaline cartilage on safranin‐O staining throughout the 26 weeks. In the deeper portion, subchondral bone and bone marrow were well remodelled. On fluoroscopic evaluation, QDs were tracked mainly in bone marrow stromata, with some signals detected in cartilage and the subchondral bone layer. We showed that the labelling of rabbit MSCs with anti‐mortalin antibody‐conjugated i‐QDs is a tolerable procedure and provides a stable fluorescence signal during the cartilage repair process for up to 26 weeks after transplantation. The results suggest that i‐MSCs did not inhibit, and indeed contributed to, the regeneration of osteochondral defects. Copyright © 2010 John Wiley & Sons, Ltd.

[1]  Tomoo Ishii,et al.  Rotating three‐dimensional dynamic culture of adult human bone marrow‐derived cells for tissue engineering of hyaline cartilage , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[2]  A. Caplan Why are MSCs therapeutic? New data: new insight , 2009, The Journal of pathology.

[3]  S. Kaul,et al.  Stable and nondisruptive in vitro/in vivo labeling of mesenchymal stem cells by internalizing quantum dots. , 2008, Human gene therapy.

[4]  P. Quesenberry,et al.  The Paradoxical Dynamism of Marrow Stem Cells: Considerations of Stem Cells, Niches, and Microvesicles , 2008, Stem Cell Reviews.

[5]  K. Takaoka,et al.  Present status of and future direction for articular cartilage repair , 2008, Journal of Bone and Mineral Metabolism.

[6]  T. Ishii,et al.  Repair of large osteochondral defects with allogeneic cartilaginous aggregates formed from bone marrow‐derived cells using RWV bioreactor , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[7]  M. Rosen,et al.  Finding Fluorescent Needles in the Cardiac Haystack: Tracking Human Mesenchymal Stem Cells Labeled with Quantum Dots for Quantitative In Vivo Three‐Dimensional Fluorescence Analysis , 2007, Stem cells.

[8]  T. Hirano,et al.  An antibody-conjugated internalizing quantum dot suitable for long-term live imaging of cells. , 2007, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[9]  Junzo Tanaka,et al.  Cartilaginous tissue formation from bone marrow cells using rotating wall vessel (RWV) bioreactor. , 2006, Biotechnology and bioengineering.

[10]  Frank Emmrich,et al.  Quantum dots for human mesenchymal stem cells labeling. A size-dependent autophagy activation. , 2006, Nano letters.

[11]  A. Caplan,et al.  Mesenchymal stem cells as trophic mediators , 2006, Journal of cellular biochemistry.

[12]  Z. Fishelson,et al.  Mortalin/GRP75 promotes release of membrane vesicles from immune attacked cells and protection from complement-mediated lysis. , 2005, International immunology.

[13]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[14]  E. Fuchs,et al.  Socializing with the Neighbors Stem Cells and Their Niche , 2004, Cell.

[15]  David E. Misek,et al.  Global Profiling of the Cell Surface Proteome of Cancer Cells Uncovers an Abundance of Proteins with Chaperone Function* , 2003, The Journal of Biological Chemistry.

[16]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[17]  S. Kaul,et al.  Identification of a novel member of mouse hsp70 family. Its association with cellular mortal phenotype. , 1993, The Journal of biological chemistry.