Characterization and osteogenic potential of equine muscle tissue– and periosteal tissue–derived mesenchymal stem cells in comparison with bone marrow– and adipose tissue–derived mesenchymal stem cells

Catherine L. Radtke Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE C1A 4P3, Canada.

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Rodolfo Nino-Fong Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE C1A 4P3, Canada.

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Blanca P. Esparza Gonzalez Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE C1A 4P3, Canada.

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Henrik Stryhn Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE C1A 4P3, Canada.

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Laurie A. McDuffee Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE C1A 4P3, Canada.

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Abstract

Objective—To characterize equine muscle tissue– and periosteal tissue–derived cells as mesenchymal stem cells (MSCs) and assess their proliferation capacity and osteogenic potential in comparison with bone marrow– and adipose tissue–derived MSCs.

Sample—Tissues from 10 equine cadavers.

Procedures—Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and adipose tissue from the left subcutaneous region. Mesenchymal stem cells were characterized on the basis of morphology, adherence to plastic, trilineage differentiation, and detection of stem cell surface markers via immunofluorescence and flow cytometry. Mesenchymal stem cells were tested for osteogenic potential with osteocalcin gene expression via real-time PCR assay. Mesenchymal stem cell cultures were counted at 24, 48, 72, and 96 hours to determine tissue-specific MSC proliferative capacity.

Results—Equine muscle tissue– and periosteal tissue–derived cells were characterized as MSCs on the basis of spindle-shaped morphology, adherence to plastic, trilineage differentiation, presence of CD44 and CD90 cell surface markers, and nearly complete absence of CD45 and CD34 cell surface markers. Muscle tissue–, periosteal tissue–, and adipose tissue–derived MSCs proliferated significantly faster than did bone marrow–derived MSCs at 72 and 96 hours.

Conclusions and Clinical Relevance—Equine muscle and periosteum are sources of MSCs. Equine muscle- and periosteal-derived MSCs have osteogenic potential comparable to that of equine adipose- and bone marrow–derived MSCs, which could make them useful for tissue engineering applications in equine medicine.

Abstract

Objective—To characterize equine muscle tissue– and periosteal tissue–derived cells as mesenchymal stem cells (MSCs) and assess their proliferation capacity and osteogenic potential in comparison with bone marrow– and adipose tissue–derived MSCs.

Sample—Tissues from 10 equine cadavers.

Procedures—Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and adipose tissue from the left subcutaneous region. Mesenchymal stem cells were characterized on the basis of morphology, adherence to plastic, trilineage differentiation, and detection of stem cell surface markers via immunofluorescence and flow cytometry. Mesenchymal stem cells were tested for osteogenic potential with osteocalcin gene expression via real-time PCR assay. Mesenchymal stem cell cultures were counted at 24, 48, 72, and 96 hours to determine tissue-specific MSC proliferative capacity.

Results—Equine muscle tissue– and periosteal tissue–derived cells were characterized as MSCs on the basis of spindle-shaped morphology, adherence to plastic, trilineage differentiation, presence of CD44 and CD90 cell surface markers, and nearly complete absence of CD45 and CD34 cell surface markers. Muscle tissue–, periosteal tissue–, and adipose tissue–derived MSCs proliferated significantly faster than did bone marrow–derived MSCs at 72 and 96 hours.

Conclusions and Clinical Relevance—Equine muscle and periosteum are sources of MSCs. Equine muscle- and periosteal-derived MSCs have osteogenic potential comparable to that of equine adipose- and bone marrow–derived MSCs, which could make them useful for tissue engineering applications in equine medicine.

Contributor Notes

This manuscript represents a portion of a thesis submitted by Dr. Radtke to the University of Prince Edward Island Department of Health Management as a partial fulfillment of the requirements for a Doctor of Philosophy degree.

Supported by Atlantic Canada Opportunities Agency and Innovation PEI.

Presented in abstract form at the 2nd Annual North American Veterinary Regenerative Medicine Conference, Lexington, Ky, June 2011.

The authors thank Dr. Glenda Wright for technical assistance.

Address correspondence to Dr. Radtke (cradtke@upei.ca).
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