The fate of transplanted xenogeneic bone marrow‐derived stem cells in rat intervertebral discs

Intervertebral disc degeneration is a major cause and a risk factor for chronic low back pain. The potential of using stem cells to treat disc degeneration has been raised. The aims of our study were to assess whether xenogeneic bone‐marrow derived stem cells could survive in a rat disc degeneration model and to determine which cell types, if any, survived and differentiated into disc‐like cells. Human bone‐marrow derived CD34+ (hematopoietic progenitor cells) and CD34− (nonhematopoietic progenitor cells, including mesenchymal stem cells) cells were isolated, fluorescent‐labeled, and injected into rat coccygeal discs. The rats were sacrificed at day 1, 10, 21, and 42. Treated discs were examined by histological and immunostaining techniques and compared to control discs. The survival of transplanted cells was further confirmed with a human nuclear specific marker. Fluorescent labeled CD34− cells were detected until day 42 in the nucleus pulposus of the injected discs. After 3 weeks these cells had differentiated into cells expressing chondrocytic phenotype (Collagen II and Sox‐9). In contrast, the fluorescent labeled CD34+ cells could not be detected after day 21. No fluorescence‐positive cells were detected in the noninjected control discs. Further, no inflammatory cells infiltrated the nucleus pulposus, even though these animals had not received immunosuppressive treatment. Our data provide evidence that transplanted human BM CD34− cells survived and differentiated within the relative immune privileged nucleus pulposus of intervertebral disc degeneration. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:374–379, 2009

[1]  I. Rasmusson Immune modulation by mesenchymal stem cells. , 2006, Experimental cell research.

[2]  Hanns-Ulrich Marschall,et al.  Mesenchymal Stem Cells for Treatment of Therapy-Resistant Graft-versus-Host Disease , 2006, Transplantation.

[3]  A. Freemont,et al.  Intervertebral Disc Cell–Mediated Mesenchymal Stem Cell Differentiation , 2006, Stem cells.

[4]  Joji Mochida,et al.  Differentiation of Mesenchymal Stem Cells Transplanted to a Rabbit Degenerative Disc Model: Potential and Limitations for Stem Cell Therapy in Disc Regeneration , 2005, Spine.

[5]  J. Lotz,et al.  The potential role of mesenchymal stem cell therapy for intervertebral disc degeneration: a critical overview. , 2005, Neurosurgical focus.

[6]  J Ratajczak,et al.  Bone marrow as a home of heterogenous populations of nonhematopoietic stem cells , 2005, Leukemia.

[7]  W. Olszewski Innate immunity processes in organ allografting--their contribution to acute and chronic rejection. , 2005, Annals of transplantation.

[8]  Anja Winter,et al.  Induction of Intervertebral Disc–Like Cells From Adult Mesenchymal Stem Cells , 2005, Stem cells.

[9]  I. Yaniv,et al.  Induction of tolerance using Fas ligand: a double-edged immunomodulator. , 2005, Blood.

[10]  Xiong Guo,et al.  Bone Mesenchymal Stem Cells Transplanted into Rabbit Intervertebral Discs Can Increase Proteoglycans , 2005, Clinical orthopaedics and related research.

[11]  Yongjun Wang,et al.  Gene expression profile of degenerated cervical intervertebral disc tissues in rats. , 2004, Chinese journal of traumatology = Zhonghua chuang shang za zhi.

[12]  P. Roughley Biology of Intervertebral Disc Aging and Degeneration: Involvement of the Extracellular Matrix , 2004, Spine.

[13]  M. Kurosaka,et al.  Fas-Ligand Expression on Nucleus Pulposus Begins in Developing Embryo , 2004, Spine.

[14]  C. Civin,et al.  Fas Ligand as a Tool for Immunosuppression and Generation of Immune Tolerance , 2004, Stem cells.

[15]  Yong-Guang Yang Application of xenogeneic stem cells for induction of transplantation tolerance: present state and future directions , 2004, Springer Seminars in Immunopathology.

[16]  M. Usuda,et al.  MACROPHAGE DEPLETION PREVENTS ANTI-GRAFT ANTIBODY PRODUCTION AND RESULTS IN LONG TERM SURVIVAL IN XENOTRANSPLANTATION , 2004, Transplantation proceedings.

[17]  Jeffrey C Lotz,et al.  Biological response of the intervertebral disc to dynamic loading. , 2004, Journal of biomechanics.

[18]  George M. Wahba,et al.  Intervertebral Disc Cell Therapy for Regeneration: Mesenchymal Stem Cell Implantation in Rat Intervertebral Discs , 2004, Annals of Biomedical Engineering.

[19]  Chulhee Choi,et al.  Fas ligand/Fas system in the brain: regulator of immune and apoptotic responses , 2004, Brain Research Reviews.

[20]  D. Bonnet Biology of human bone marrow stem cells , 2003, Clinical and Experimental Medicine.

[21]  Safdar N. Khan,et al.  The molecular basis of intervertebral disk degeneration. , 2003, The Orthopedic clinics of North America.

[22]  M. Lübbert,et al.  CD34− Hematopoietic Stem Cells: Current Concepts and Controversies , 2003, Stem cells.

[23]  S. Roberts,et al.  Disc morphology in health and disease. , 2002, Biochemical Society transactions.

[24]  M. Kurosaka,et al.  Fas Ligand Exists on Intervertebral Disc Cells: A Potential Molecular Mechanism for Immune Privilege of the Disc , 2002, Spine.

[25]  M. Bhatia,et al.  Characterization of retroviral gene transfer into highly purified human CD34(-) cells with primitive hematopoietic capacity. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[26]  A. Freemont,et al.  Current understanding of cellular and molecular events in intervertebral disc degeneration: implications for therapy , 2002, The Journal of pathology.

[27]  R. Colvin,et al.  Tolerance, Mixed Chimerism, and Chronic Transplant Arteriopathy1 2 , 2001, The Journal of Immunology.

[28]  Jong-Beom Park,et al.  Expression of Fas Ligand and Apoptosis of Disc Cells in Herniated Lumbar Disc Tissue , 2001, Spine.

[29]  D. Krause,et al.  Hematopoietic Stem Cells Can Be CD34+ or CD34- , 2001, Leukemia & lymphoma.

[30]  R. Fessler,et al.  Molecular Biology of Degenerative Disc Disease , 2000, Neurosurgery.

[31]  F. Cammisa,et al.  Current concepts in intervertebral disc restoration. , 2000, The Orthopedic clinics of North America.

[32]  J. Nolta,et al.  CD34: To select or not to select? That is the question , 2000, Leukemia.

[33]  S. Nagata,et al.  Fas ligand-induced apoptosis. , 1999, Annual review of genetics.

[34]  M. Ogawa,et al.  Engraftment and Multilineage Expression of Human Bone Marrow CD34− Cells In Vivo a , 1999, Annals of the New York Academy of Sciences.

[35]  M. Ogawa,et al.  Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. , 1998, Experimental hematology.

[36]  K. Okumura,et al.  CD95 ligand in graft rejection , 1996, Nature.

[37]  D. Green,et al.  Fas Ligand-Induced Apoptosis as a Mechanism of Immune Privilege , 1995, Science.

[38]  K. Ando,et al.  Regenerative effects of transplanting mesenchymal stem cells embedded in atelocollagen to the degenerated intervertebral disc. , 2006, Biomaterials.

[39]  J. Mosca,et al.  T cell responses to allogeneic human mesenchymal stem cells: immunogenicity, tolerance, and suppression. , 2005, Journal of biomedical science.

[40]  H. Brisby,et al.  Cell therapy for disc degeneration--potentials and pitfalls. , 2004, The Orthopedic clinics of North America.

[41]  Kevin McIntosh,et al.  Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. , 2002, Experimental hematology.

[42]  J. Dick,et al.  Distinct classes of human stem cells that differ in proliferative and self-renewal potential , 2001, Nature Immunology.

[43]  M. Fackler,et al.  CD34: structure, biology, and clinical utility. , 1996, Blood.