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Cartilage Engineering from Ovine Umbilical Cord Blood Mesenchymal Progenitor Cells

^a Children’s Hospital Boston,
^b Massachusetts General Hospital,
^c Harvard Medical School Center for Minimally Invasive Surgery, Boston, Massachusetts, USA

Key Words. Cord blood cells • Differentiation • Fetal stem cells • In vitro differentiation • Stem/progenitor cell • Stromal cells • TGF-ß1 • Tissue regeneration

Correspondence: Dario O. Fauza, M.D., Children’s Hospital Boston, Department of Surgery, Fegan 3, 300 Longwood Avenue, Boston, Massachusetts 02115, USA. Telephone: 617-919-2966; Fax: 617-730-0910; e-mail: dario.fauza{at}childrens.harvard.edu

We aimed to determine whether three-dimensional (3D) cartilage could be engineered from umbilical cord blood (CB) cells and compare it with both engineered fetal cartilage and native tissue. Ovine mesenchymal progenitor cells were isolated from CB samples (n = 4) harvested at 80–120 days of gestation by low-density fractionation, expanded, and seeded onto polyglycolic acid scaffolds. Constructs (n = 28) were maintained in a rotating bioreactor with serum-free medium supplemented with transforming growth factor-ß1 for 4–12 weeks. Similar constructs seeded with fetal chondrocytes (n = 13) were cultured in parallel for 8 weeks. All specimens were analyzed and compared with native fetal cartilage samples (n = 10). Statistical analysis was by analysis of variance and Student’s t-test (p < .01). At 12 weeks, CB constructs exhibited chondrogenic differentiation by both standard and matrix-specific staining. In the CB constructs, there was a significant time-dependent increase in extracellular matrix levels of glycosaminoglycans (GAGs) and type-II collagen (C-II) but not of elastin (EL). Fetal chondrocyte and CB constructs had similar GAG and C-II contents, but CB constructs had less EL. Compared with both hyaline and elastic native fetal cartilage, C-II and EL levels were, respectively, similar and lower in the CB constructs, which had correspondingly lower and similar GAG levels than native hyaline and elastic fetal cartilage. We conclude that CB mesenchymal progenitor cells can be successfully used for the engineering of 3D cartilaginous tissue in vitro, displaying select histological and functional properties of both native and engineered fetal cartilage. Cartilage engineered from CB may prove useful for the treatment of select congenital anomalies.

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