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Comparison of the osteogenic potential of equine mesenchymal stem cells from bone marrow, adipose tissue, umbilical cord blood, and umbilical cord tissue

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  • 1 Departments of Anatomy, Physiology, and Cell Biology, University of California-Davis, Davis, CA 95616.
  • | 2 Departments of Anatomy, Physiology, and Cell Biology, University of California-Davis, Davis, CA 95616.
  • | 3 Departments of Anatomy, Physiology, and Cell Biology, University of California-Davis, Davis, CA 95616.
  • | 4 School of Veterinary Medicine, and Department of Biomedical Engineering, College of Engineering, University of California-Davis, Davis, CA 95616.
  • | 5 Pathology, Microbiology, and Immunology, University of California-Davis, Davis, CA 95616.
  • | 6 Center for Equine Health, University of California-Davis, Davis, CA 95616.
  • | 7 Surgical and Radiological Sciences, University of California-Davis, Davis, CA 95616.
  • | 8 School of Veterinary Medicine, and Department of Biomedical Engineering, College of Engineering, University of California-Davis, Davis, CA 95616.
  • | 9 Pathology, Microbiology, and Immunology, University of California-Davis, Davis, CA 95616.
  • | 10 Departments of Anatomy, Physiology, and Cell Biology, University of California-Davis, Davis, CA 95616.

Abstract

Objective—To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells.

Animals—10 adult Quarter Horses and 8 newborn Thoroughbred foals.

Procedures—eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated.

Results—BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP activity and calcium deposition. There was no significant difference in BM-eMSC yield from the tuber coxae or sternum, but yield was significantly greater from the first 60-mL aspirate than from subsequent aspirates. The BM-eMSC expansion rate was significantly higher when cells were cultured in fetal bovine serum instead of autologous serum (AS).

Conclusions and Clinical Relevance—eMSCs from BM possessed the highest in vitro osteogenic potential; eMSCs from adipose tissue also had robust osteogenic potential. The tuber coxae and the sternum were viable sources of BM-eMSCs in yearlings, and 60 mL of BM aspirate was sufficient for culture and expansion. Expanding BM-eMSCs in AS to avoid potential immunologic reactions decreased the total yield because BM-eMSCs grew significantly slower in AS than in fetal bovine serum. Additional studies are needed to determine optimal ex vivo eMSC culture and expansion conditions, including the timing and use of growth factor—supplemented AS. (Am J Vet Res 2010;71:1237-1245)

Abstract

Objective—To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells.

Animals—10 adult Quarter Horses and 8 newborn Thoroughbred foals.

Procedures—eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated.

Results—BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP activity and calcium deposition. There was no significant difference in BM-eMSC yield from the tuber coxae or sternum, but yield was significantly greater from the first 60-mL aspirate than from subsequent aspirates. The BM-eMSC expansion rate was significantly higher when cells were cultured in fetal bovine serum instead of autologous serum (AS).

Conclusions and Clinical Relevance—eMSCs from BM possessed the highest in vitro osteogenic potential; eMSCs from adipose tissue also had robust osteogenic potential. The tuber coxae and the sternum were viable sources of BM-eMSCs in yearlings, and 60 mL of BM aspirate was sufficient for culture and expansion. Expanding BM-eMSCs in AS to avoid potential immunologic reactions decreased the total yield because BM-eMSCs grew significantly slower in AS than in fetal bovine serum. Additional studies are needed to determine optimal ex vivo eMSC culture and expansion conditions, including the timing and use of growth factor—supplemented AS. (Am J Vet Res 2010;71:1237-1245)

Contributor Notes

Supported by a private grant from Dick and Carolyn Randall and the Harriet E. Pfleger Foundation.

The authors thank Fred Librach for equine autologous serum collection as well as Naomi Walker, Danielle Carrade, Michael Friedman, and Sivan Oyserman for the isolation and expansion of umbilical cord tissue and umbilical cord blood mesenchymal stem cells.

Address correspondence to Dr. Yellowley (cyellowley@ucdavis.edu).