J Vet Sci.  2013 Sep;14(3):367-371. 10.4142/jvs.2013.14.3.367.

Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood

• Kang JG
• Park SB
• Seo MS
• Kim HS
• Chae JS
• Kang KS

• KMID: 1705569
• DOI: http://doi.org/10.4142/jvs.2013.14.3.367

Abstract

Tendinitis of the superficial digital flexor tendon (SDFT) is a significant cause of lameness in horses; however, recent studies have shown that stem cells could be useful in veterinary regenerative medicine. Therefore, we isolated and characterized equine umbilical cord blood mesenchymal stem cells (eUCB-MSCs) from equine umbilical cord blood obtained from thoroughbred mares during the foaling period. Horses that had tendinitis of the SDFT were treated with eUCB-MSCs to confirm the therapeutic effect. After eUCB-MSCs transplantation, the core lesion in the SDFT was found to decrease. These results suggest that transplantation using eUCB-MSCs could be another source of cell treatment.

Keyword

cell transplantation; equine; mesenchymal stem cells; umbilical cord blood

MeSH Terms

Animals
*Cord Blood Stem Cell Transplantation/veterinary
Horse Diseases/*surgery
Horses
Male
Tendinopathy/surgery/*veterinary

Figure

• 

  Fig. 1 Primary culture, cumulative population doubling level (CPDL), and differentiation of equine umbilical cord blood mesenchymal stem cells (eUCB-MSCs). (A) Phase contrast images of eUCB-MSCs. Cell morphology showed a spindle and fibroblast-like structure that was similar to that of human mesenchymal stem cells. (B) Cell growth curve of eUCB-MSCs. CPDL was measured from passages 3 to 18. (C~F) Osteogenic differentiation. Alizarin Red S staining was conducted after 3 weeks of osteogenic induction. Osteogenic differentiated cells (C, D) were grown in osteogenic induction medium and differentiated cells stained strongly with Alizarin Red S. Control cells (E, F) were grown in normal low glucose DMEM with 10% FBS and showed no staining with Alizarin Red S. (G~J) Adipogenic differentiation. Oil Red O staining was conducted after 3 weeks of adipogenic induction. Adipogenic differentiated cells (G, H) were grown in adipogenic induction medium and differentiated cells were stained with Oil Red O. The black arrows indicate stained red fat droplets. Control cells (I, J) were grown in normal low glucose DMEM with 10% FBS and no staining with Oil Red O was observed. (K, L) Chondrogenic differentiation. After 3 weeks of chondrogenic induction, pellets aggregated into a round shape. (K) Pictures of the round shaped chondrogenic pellet. A pellet formed at the bottom of the 15 mL polypropylene tube. The white arrow indicates a pellet. (L) Toluidine blue staining of chondrogenic pellets. Scale bars = 50 µm (A, D, F, H and J), and 100 µm (L).

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  Fig. 2 Ultrasonographic images of a core lesion in a superficial digital flexor tendon (white dashed circle). Before eUCB-MSCs injection (A), 1 month after MSCs injection (B), and 3 months after MSCs injection (C). Note the rapid filling-in of the lesion and the absence of apparent adverse effects.

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