Evaluation of equine peripheral blood apheresis product, bone marrow, and adipose tissue as sources of mesenchymal stem cells and their differentation potential.

OBJECTIVE To evaluate effects of apheresis on mesenchymal stem cells (MSCs) and compare those MSCs with MSCs obtained from adipose tissue or bone marrow (BM). SAMPLE POPULATION Samples obtained from 6 adult horses. PROCEDURES Samples of blood from a peripheral vein, adipose tissue, and BM aspirate were obtained from each horse. Samples were processed via apheresis of blood and techniques reported elsewhere for adipose tissue and BM. Cultures were maintained until adherence and subsequently were subjected to differentiation protocols to evaluate adipogenic, osteoblastogenic, and chondrogenic potential. RESULTS Apheresis product had a significantly higher mononuclear percentage, higher platelet count, and lower RBC count, compared with values for peripheral blood. No cell adherence to the tissue culture plates was detected for the apheresis product. Adherence was detected for 6 of 6 adipose-derived and 4 of 6 BM-derived samples. Variations in efficiency were detected for differentiation of adipose- and BM-derived cells into adipocytes, chondrocytes, and osteoblasts. CONCLUSIONS AND CLINICAL RELEVANCE Apheresis was able to concentrate mononuclear cells and reduce RBC contamination. However, the apheresis product was unable to adhere to the tissue culture plates. In matched horses, adipose- and BM-derived MSCs were capable of producing lipids, glycosaminoglycan, and mineral. The BM was vastly superior to adipose tissue as a source of MSCs with osteoblastogenic potential in matched horses. Additional studies will be necessary to optimize apheresis techniques for horses before peripheral blood can be considered a suitable source for multipotential cells for use in cell-based treatments.

[1]  C. Kawcak,et al.  Evaluation of adipose‐derived stromal vascular fraction or bone marrow‐derived mesenchymal stem cells for treatment of osteoarthritis , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[2]  Marjolein C H van der Meulen,et al.  Mesenchymal stem cells and insulin‐like growth factor‐I gene‐enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  A. Grodzinsky,et al.  Evaluation of adult equine bone marrow‐ and adipose‐derived progenitor cell chondrogenesis in hydrogel cultures , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  P. Mainil-Varlet,et al.  Multilineage differentiation potential of equine blood-derived fibroblast-like cells. , 2008, Differentiation; research in biological diversity.

[5]  Roger K. Smith Mesenchymal stem cell therapy for equine tendinopathy , 2008, Disability and rehabilitation.

[6]  K. Addicks,et al.  Isolation and characterization of bone marrow-derived equine mesenchymal stem cells. , 2007, American journal of veterinary research.

[7]  J. Bartela,et al.  Hyaluronic acid and autologous synovial fluid induce chondrogenic differentiation of equine mesenchymal stem cells : a preliminary study , 2007 .

[8]  N. Kobayashi,et al.  Stem cell-derived hepatocytes , 2006 .

[9]  Mandi J. Lopez,et al.  Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity. , 2006, Veterinary surgery : VS.

[10]  L. Gepstein,et al.  Stem cells for myocardial repair , 2006 .

[11]  M. Conese,et al.  Stem cells and cystic fibrosis. , 2006, Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society.

[12]  P. Mainil-Varlet,et al.  Equine Peripheral Blood‐Derived Progenitors in Comparison to Bone Marrow‐Derived Mesenchymal Stem Cells , 2006, Stem cells.

[13]  L. Zangi,et al.  Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads , 2006, Bone Marrow Transplantation.

[14]  R. Burt,et al.  Nonmyeloablative hematopoietic stem cell transplantation for systemic lupus erythematosus. , 2006, JAMA.

[15]  Wei Liu,et al.  Bone reconstruction with bone marrow stromal cells. , 2006, Methods in enzymology.

[16]  L. Lagneaux,et al.  Mesenchymal Stem Cells Derived from CD133‐Positive Cells in Mobilized Peripheral Blood and Cord Blood: Proliferation, Oct4 Expression, and Plasticity , 2005, Stem cells.

[17]  G. Schuurhuis,et al.  A single-step colony-forming unit assay for unseparated mobilized peripheral blood, cord blood, and bone marrow. , 2001, Journal of hematotherapy & stem cell research.

[18]  R. Baron,et al.  Opposite effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on osteoblast differentiation. , 2001, Bone.

[19]  K. Satomura,et al.  Circulating Skeletal Stem Cells , 2001, The Journal of cell biology.

[20]  G. Schuurhuis,et al.  Research Report A Single-Step Colony-Forming Unit Assay for Unseparated Mobilized Peripheral Blood, Cord Blood, and Bone Marrow , 2001 .

[21]  Jill Moss,et al.  Mesenchymal precursor cells in the blood of normal individuals , 2000, Arthritis Research & Therapy.

[22]  E. Weissinger,et al.  Evidence of Peripheral Blood‐Derived, Plastic‐Adherent CD34−/low Hematopoietic Stem Cell Clones with Mesenchymal Stem Cell Characteristics , 2000, Stem cells.

[23]  J. Williams,et al.  Isolation and chondrocytic differentiation of equine bone marrow-derived mesenchymal stem cells. , 1998, American journal of veterinary research.