Effect of fibroblast growth factor-2 on equine mesenchymal stem cell monolayer expansion and chondrogenesis

Stewart, Alban; Byron, Christopher R.; Pondenis, Holly C.; Stewart, Margaret M.

doi:10.2460/ajvr.68.9.941

Cited by 89 publications

(74 citation statements)

References 32 publications

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“…After normalizing matrix synthesis to cell number, the difference was not significant; much of the difference in matrix synthesis is attributable to the differences in cell attachment and survival over time. These results were similar to those of previous studies 44,45 of matrix synthesis by equine bone marrow-derived MSCs, in which increased synthesis reflected increases in cell numbers rather than per-cell changes. Nevertheless, this outcome is relevant to therapeutic applications because colonization of the tendon matrix and persistence at the site of administration would likely influence the clinical efficacy of these cells for treatment of horses with tendon injuries.…”

Section: Discussionsupporting

confidence: 82%

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Comparison of equine tendon-, muscle-, and bone marrow–derived cells cultured on tendon matrix

Stewart

Barrett

Byron

et al. 2009

ajvr

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Objective-To compare viability and biosynthetic capacities of cells isolated from equine tendon, muscle, and bone marrow grown on autogenous tendon matrix. Sample Population-Cells from 4 young adult horses. Procedures-Cells were isolated, expanded, and cultured on autogenous cell-free tendon matrix for 7 days. Samples were analyzed for cell viability, proteoglycan synthesis, collagen synthesis, and mRNA expression of collagen type I, collagen type III, and cartilage oligomeric matrix protein (COMP). Results-Tendon-and muscle-derived cells required less time to reach confluence (approx 2 weeks) than did bone marrow-derived cells (approx 3 to 4 weeks); there were fewer bone marrow-derived cells at confluence than the other 2 cell types. More tendon-and musclederived cells were attached to matrices after 7 days than were bone marrow-derived cells. Collagen and proteoglycan synthesis by tendon-and muscle-derived cells was significantly greater than synthesis by bone marrow-derived cells. On a per-cell basis, tendon-derived cells had more collagen synthesis, although this was not significant. Collagen type I mRNA expression was similar among groups. Tendon-derived cells expressed the highest amounts of collagen type III and COMP mRNAs, although the difference for COMP was not significant. Conclusions and Clinical Relevance-Tendon-and muscle-derived cells yielded greater cell culture numbers in shorter time and, on a per-cell basis, had comparable biosynthetic assays to bone marrow-derived cells. More in vitro experiments with higher numbers may determine whether tendon-derived cells are a useful resource for tendon healing. (Am J Vet Res 2009;70:750-757)

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Section: Discussionsupporting

confidence: 82%

“…On the basis of these data, a minimum of 12 horses would be necessary to provide sufficient power (0.8) for these analyses. However, in our experience, marked interdonor variability is a biological reality of mesenchymal cell isolation and in vitro activity 18,44,45 and is likely to be reflected in in vivo therapeutic efficacies.…”

Section: Discussionmentioning

confidence: 99%

Comparison of equine tendon-, muscle-, and bone marrow–derived cells cultured on tendon matrix

Stewart

Barrett

Byron

et al. 2009

ajvr

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“…The time required to generate clinically useful cell numbers could be reduced by collecting larger volumes of synovial fluid and/or by aspirating fluid from several joints, given that arthrocentesis is a minimally invasive procedure, compared to bone marrow or adipose tissue harvesting. In vitro expansion of SF-CPs can also be substantially accelerated by fibroblast growth factor-2, reducing expansion times by more than 50%, without compromising subsequent chondrogenesis [21], as with bone marrow MSCs [30]. Alternatively, endogenous SF-CPs could be directly recruited to sites of cartilage damage to improve healing, by modulating chemokine concentrations at sites of injury [2,31,32] and utilizing the intrinsic chondrogenic properties of synovial fluid [33].…”

Section: Discussionmentioning

confidence: 99%

Phenotypic characterization of equine synovial fluid-derived chondroprogenitor cells

Chen¹,

Bianchessi²,

Pondenis³

et al. 2016

Stem Cell Biol Res

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Background: Progenitor cells exist in most tissues and body fluids. Synovial fluid chondroprogenitor cells have been described in several species; however, the specific phenotypic characteristics of these cells have not been defined. This study addressed the impacts of joint location and donor age variation on synovial fluid chondroprogenitor numbers, and determined whether synovial fluid chondroprogenitors express hypertrophic characteristics or a non-endochondral phenotype in chondrogenic culture. Methods: Synovial fluid was aspirated from the joints of healthy adult horses, and cells in the aspirates were expanded through two monolayer passages. The number of colony-forming units (CFU) in each aspirate was assessed after 14 days. Population doublings (PD) and PD times were calculated during the subsequent two passages. Passage 3 cells were used to determine osteogenic and adipogenic capacities, or were transferred to pellet cultures in chondrogenic medium containing TGF-β1 or BMP-2. Bone marrowderived MSCs and primary articular chondrocytes were cultured under similar conditions to provide reference values for chondrogenesis. Chondrogenesis was assessed by measuring collagen type II protein and sulfated glycosaminoglycan secretion, expression of chondrogenic mRNAs, and induction of ALP activity. Results: CFU counts varied considerably, but the majority of aspirates contained <50 CFU/ml. PDs were consistently between 2.0 and 3.0 during both passages. PD times varied from 0.2-0.4 days. Proliferation was not significantly influenced by anatomical location or donor age. Equine synovial fluid progenitors expressed osteogenic, adipogenic and chondrogenic phenotypes under appropriate conditions. Under chondrogenic conditions, synovial fluid cells increased collagen and aggrecan mRNA expression to levels comparable to bone marrow MSCs, although collagen type II protein secretion was less than half that of articular chondrocytes. In contrast to bone marrow MSCs, synovial fluid chondroprogenitors did not express collagen type X or increase ALP activity in response to TGF-β1 or BMP-2. Consistent with these observations, Runx2 and Mef2C expression in synovial fluid chondroprogenitors was 20-40 fold lower than in bone marrow MSCs. Conclusions: Synovial fluid chondroprogenitors are capable of robust non-hypertrophic chondrogenesis, whether stimulated by TGF-β1 or by BMP-2. These results indicate that synovial fluid cells are phenotypically suitable for articular cartilage repair/regeneration, although strategies to accelerate cell proliferation and synthesize a functional cartilage matrix in vivo will be required for clinically feasible cellbased therapies.

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“…Since FGF2 and IGF1 have been reported to be growth factors which enhance the proliferation or differentiation of chondrocytes and their progenitors, it could be expected that high expression of these factors contributes to repair of a cartilage defect. 24,25 However, FGF2 may also be disadvantageous for cartilage formation because of its proangiogenic and pro-osteogenic potential. Miyakoshi et al demonstrated that intra-articular administration of FGF2 induced inflammation and osteophyte formation.…”

Section: Figmentioning

confidence: 99%

Repair Mechanism of Osteochondral Defect Promoted by Bioengineered Chondrocyte Sheet

Shimizu

Kamei

Adachi

et al. 2015

Tissue Engineering Part A

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Cell sheet engineering has developed as a remarkable method for cell transplantation. In the field of cartilage regeneration, several studies previously reported that cartilage defects could be regenerated by transplantation of a chondrocyte sheet using cell sheet engineering. However, it remains unclear how such a thin cell sheet could repair a deep cartilage defect. We, therefore, focused on the mechanism of cartilage repair using cell sheet engineering in this study. Chondrocyte sheets and synovial cell sheets were fabricated using cell sheet engineering, and these allogenic cell sheets were transplanted to cover an osteochondral defect in a rat model. Macroscopic and histological evaluation was performed at 4 and 12 weeks after transplantation. Analysis of the gene expression of each cell sheet and of the regenerated tissue at 1 week after transplantation was performed. In addition, green fluorescent protein (GFP) transgenic rats were used as donors (transplanted chondrocyte sheets) or recipients (osteochondral defect models) to identify the cell origin of regenerated cartilage.Cartilage repair was significantly better in the group implanted with a chondrocyte sheet than in that with a synovial cell sheet. The results of gene expression analysis suggest that the possible factor contributing to cartilage repair might be TGFb1. Cell tracking experiments using GFP transgenic rats showed that the regenerated cartilage was largely composed of cells derived from the transplanted chondrocyte sheets.

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Effect of fibroblast growth factor-2 on equine mesenchymal stem cell monolayer expansion and chondrogenesis

Abstract: FGF-2 treatment of bone marrow-derived MSC monolayers enhanced subsequent chondrogenic differentiation in a 3-dimensional culture. This result is important for tissue engineering strategies dependent on MSC expansion for cartilage repair.

Cited by 89 publications

References 32 publications

Comparison of equine tendon-, muscle-, and bone marrow–derived cells cultured on tendon matrix

Comparison of equine tendon-, muscle-, and bone marrow–derived cells cultured on tendon matrix

Phenotypic characterization of equine synovial fluid-derived chondroprogenitor cells

Repair Mechanism of Osteochondral Defect Promoted by Bioengineered Chondrocyte Sheet

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