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Abstract

Objective—To determine the morphologic and phenotypic effects of transforming growth factor β1 (TGF-β1) on cultured equine mesenchymal stem cells (MSC) and articular chondrocytes.

Sample Population—Bone marrow aspirates and articular cartilage samples from a 2-year-old and two 8- month-old horses.

Procedure—After initial isolation and culture, MSC and chondrocytes were cultured in Ham's F-12 medium supplemented with TGF-β1 at a concentration of 0, 1, 5, or 10 ng/ml. Medium was exchanged on day 2, and cells were harvested on day 4. Medium was assayed for proteoglycan (PG) content. Total RNA was isolated from cell cultures, and expression of aggrecan, decrin, collagen type-I, and collagen type-II mRNA was assessed by means of Northern blot analyses. Cell cultures were stained with H&E or toluidine blue and examined histologically. Additional cultures were examined after immunohistochemical staining for type-I and -II collagen.

Results—MSC cultures exposed to TGF-β1 had an increased cellular density with cell layering and nodule formation that was most pronounced in cultures treated with 5 ng of TGF-β1/ml. Expression of collagen type-II mRNA in MSC cultures exposed to 5 ng of TGF- β1/ml was 1.7 times expression in control cultures, and expression of collagen type-I mRNA was 2.8 times expression in control cultures. Treatment of MSC with TGF-β1 led to dose-related increases in area and intensity of type-II collagen immunoreaction.

Conclusion—Results suggest that TGF-β1 enhances chondrogenic differentiation of bone marrow-derived MSC in a dose-dependent manner. (Am J Vet Res 2000;61:1003–1010)

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in American Journal of Veterinary Research

In the report “Evaluation of autologous bone marrow–derived mesenchymal stem cells on renal regeneration after experimentally induced acute kidney injury in dogs” ( Am J Vet Res 2016;77:208–217), the address for Dr. Jae-Ik Han is incorrect

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in American Journal of Veterinary Research

Abstract

Objective—To examine age-related efficacy of bone morphogenetic protein (BMP)-2, ascorbate, and dexamethasone as osteogenic inducers in canine marrow-derived stromal cells (MSCs).

Sample Population—Samples of femoral bone marrow obtained from 15 skeletally immature (< 1 year old) and 4 skeletally mature (> 1.5 years old) dogs.

Procedure—First-passage canine MSC cultures were treated with 100 µg of ascorbate phosphate/mL, 10–7M dexamethasone, 100 ng of BMP-2/mL, or a combination of these osteoinducers. On day 6, cultures were harvested for quantitation of alkaline phosphatase (ALP) activity and isolation of RNA to prepare cDNA for real-time polymerase chain reaction analyses of osteoblast markers.

Results—Early markers of osteogenesis were induced in canine MSCs by BMP-2 but not dexamethasone. In young dogs, the combination of BMP- 2 and ascorbate yielded the highest ALP mRNA concentrations and activity. This combination also induced significant increases in mRNA for osteopontin and runt-domain transcription factor 2. In comparison to MSCs from immature dogs, those from mature dogs had diminished ALP activity in response to BMP and ascorbate. Results for cultures treated with 3,4-dehydroproline suggested that ascorbateinduced production of extracellular matrix was important for maximal BMP-2 response in canine MSCs.

Conclusions and Clinical Relevance—BMP-2 was capable of inducing markers of osteogenesis in shortterm cultures of canine MSCs. In MSCs obtained from skeletally immature dogs, ascorbate was required for maximal effects of BMP. These results define optimal conditions for stem cell osteogenesis in dogs and will facilitate development of stem cell–based treatments for dogs with fractures. (Am J Vet Res 2005;66:1729–1737)

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in American Journal of Veterinary Research

limited by religious or ethical issues, their potential for tissue regeneration is being actively researched. Mesenchymal stem cells are widely used for several clinical applications including articular cartilage repair. 3 Autologous or allogenic MSCs

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in American Journal of Veterinary Research

platelet-derived growth factor, transforming growth factor-β, fibroblastic growth factor, vascular endothelial growth factor, insulin-like growth factor-I, and epidermal growth factor. 3 Mesenchymal stem cells may be isolated and cultured from equine bone

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in American Journal of Veterinary Research
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. Subconjunctival bone marrow-derived mesenchymal stem cell therapy as a novel treatment alternative for equine immune-mediated keratitis: a case series . Vet Ophthalmol . 2019 ; 22 ( 5 ): 674 – 682 . doi: 10.1111/vop.12641 30715781 13. Schnabel LV

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in American Journal of Veterinary Research

Bone marrow–derived mesenchymal stem cells are pluripotent and may differentiate into many mesenchymal lineages, including muscle, bone, cartilage, tendon, and ligament, and may be carrier cells for ex vivo gene transfer. 34–39 Localized delivery of

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in American Journal of Veterinary Research

F eline mesenchymal stem cells (MSCs; sometimes referred to as mesenchymal stromal cells) were first mentioned in the literature in 2002. 1 Significant interest has been placed on the potential of MSC and related animal cell and tissue

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in Journal of the American Veterinary Medical Association

– 560 . doi: 10.5966/sctm.2015-0261 10. Carrade Holt DD , Wood JA , Granick JL , Walker NJ , Clark KC , Borjesson DL . Equine mesenchymal stem cells inhibit T cell proliferation through different mechanisms depending on tissue source

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in Journal of the American Veterinary Medical Association

.3109/17453674.2010.539498 10.3109/17453674.2010.539498 25709479 3. Leroux L , Descamps B , Tojais NF , Hypoxia preconditioned mesenchymal stem cells improve vascular and skeletal muscle fiber regeneration after ischemia through a Wnt4-dependent pathway . Mol

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in Journal of the American Veterinary Medical Association