Responses of equine tendon- and bone marrow-derived cells to monolayer expansion with fibroblast growth factor-2 and sequential culture with pulverized tendon and insulin-like growth factor-I.
Abstract: To compare in vitro expansion of equine tendon- and bone marrow-derived cells with fibroblast growth factor-2 (FGF-2) supplementation and sequential matrix synthesis with pulverized tendon and insulin-like growth factor-I (IGF-I). Methods: Cells from 6 young adult horses. Methods: Progenitor cells were expanded in monolayers with FGF-2, followed by culture with autogenous acellular pulverized tendon and IGF-I for 7 days. Initial cell isolation and subsequent monolayer proliferation were assessed. In pulverized tendon cultures, cell viability and expression of collagen types I and III and cartilage oligomeric matrix protein (COMP) mRNAs were assessed. Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period. Results: Monolayer expansion with FGF-2 significantly increased the mean ± SE number of tendon-derived cells (15.3 ± 2.6 × 10(6)), compared with bone marrow-derived cells (5.8 ± 1.8 × 10(6)). Overall, increases in collagen type III and COMP mRNAs were seen in tendon-derived cells, compared with results for bone marrow-derived cells. After IGF-I supplementation, increases in collagen type I and type III mRNA expression were seen in bone marrow-derived cells, compared with results for unsupplemented control cells. Insulin-like growth factor-I significantly increased collagen synthesis of bone marrow-derived cells. Monolayer expansion with FGF-2 followed by IGF-I supplementation significantly increased glycosaminoglycan synthesis in tendon-derived cells. Conclusions: Tendon-derived cells had increased cell numbers and matrix synthesis after monolayer expansion with FGF-2, compared with results for bone marrow-derived cells. In vivo experiments with FGF-2-expanded tendon-derived cells are warranted to evaluate effects on tendon healing.
Publication Date: 2011-12-30 PubMed ID: 22204303DOI: 10.2460/ajvr.73.1.162Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper discusses an experiment comparing the impact of fibroblast growth factor-2 (FGF-2) on equine tendon and bone marrow-derived cells. It further explores the subsequent effects of insulin-like growth factor-I (IGF-I) and pulverized tendon on these cells. The results indicate that tendon-derived cells showed a higher increase in cell numbers and matrix synthesis when compared to bone marrow-derived cells.
Methodology
- The study was conducted using cells extracted from six young adult horses.
- Progenitor cells were grown in monolayers with FGF-2, followed by culture with autogenous acellular pulverized tendon and IGF-I for a week.
- The success of the initial cell isolation and subsequent proliferation in the monolayer was considered.
- In pulverized tendon cultures, researchers checked for cell viability, and expression of collagen types I and III and cartilage oligomeric matrix protein (COMP) mRNAs.
- Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period.
Results
- Monolayer expansion with FGF-2 significantly boosted the average number of tendon-derived cells compared to bone marrow-derived cells.
- Increases in collagen type III and COMP mRNAs were observed in tendon-derived cells in contrast to bone marrow-derived cells.
- After IGF-I supplementation, there was a noticeable increase in collagen type I and type III mRNA expression in bone marrow-derived cells in comparison to unsupplemented control cells.
- IGF-I considerably increased the collagen synthesis in bone marrow-derived cells.
- Monolayer expansion with FGF-2 followed by IGF-I supplementation significantly bumped up the glycosaminoglycan synthesis in tendon-derived cells.
Conclusions
- The results underscored that tendon-derived cells demonstrated a bigger surge in cell numbers and in matrix synthesis following monolayer expansion with FGF-2 compared to the bone marrow-derived cells.
- Given these benefits of using FGF-2, the researchers suggest the importance of carrying out in vivo experiments using FGF-2-expanded tendon-derived cells to further evaluate their effects on tendon healing.
Cite This Article
APA
Durgam SS, Stewart AA, Pondenis HC, Yates AC, Evans RB, Stewart MC.
(2011).
Responses of equine tendon- and bone marrow-derived cells to monolayer expansion with fibroblast growth factor-2 and sequential culture with pulverized tendon and insulin-like growth factor-I.
Am J Vet Res, 73(1), 162-170.
https://doi.org/10.2460/ajvr.73.1.162 Publication
Researcher Affiliations
- Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61802, USA.
MeSH Terms
- Animals
- Blotting, Northern / veterinary
- Bone Marrow Cells / cytology
- Bone Marrow Cells / drug effects
- Bone Marrow Cells / metabolism
- Bone Marrow Cells / physiology
- Cell Culture Techniques / methods
- Cell Culture Techniques / veterinary
- Collagen Type I / biosynthesis
- Collagen Type III / biosynthesis
- Extracellular Matrix / drug effects
- Extracellular Matrix / metabolism
- Extracellular Matrix Proteins / biosynthesis
- Fibroblast Growth Factor 2 / pharmacology
- Gene Expression Regulation
- Glycoproteins / biosynthesis
- Glycosaminoglycans / biosynthesis
- Horses / metabolism
- Insulin-Like Growth Factor I / pharmacology
- Matrilin Proteins
- RNA, Messenger / metabolism
- Real-Time Polymerase Chain Reaction / veterinary
- Tendons / cytology
- Tendons / drug effects
- Tendons / growth & development
- Tendons / metabolism
Citations
This article has been cited 8 times.- Roberts JH, Halper J. Growth Factor Roles in Soft Tissue Physiology and Pathophysiology.. Adv Exp Med Biol 2021;1348:139-159.
- Sullivan SN, Altmann NN, Brokken MT, Durgam SS. In vitro Effects of Methylprednisolone Acetate on Equine Deep Digital Flexor Tendon-Derived Cells.. Front Vet Sci 2020;7:486.
- Durgam SS, Altmann NN, Coughlin HE, Rollins A, Hostnik LD. Insulin Enhances the In Vitro Osteogenic Capacity of Flexor Tendon-Derived Progenitor Cells.. Stem Cells Int 2019;2019:1602751.
- Su Y, Denbeigh JM, Camilleri ET, Riester SM, Parry JA, Wagner ER, Yaszemski MJ, Dietz AB, Cool SM, van Wijnen AJ, Kakar S. Extracellular matrix protein production in human adipose-derived mesenchymal stem cells on three-dimensional polycaprolactone (PCL) scaffolds responds to GDF5 or FGF2.. Gene Rep 2018 Mar;10:149-156.
- Burk J, Plenge A, Brehm W, Heller S, Pfeiffer B, Kasper C. Induction of Tenogenic Differentiation Mediated by Extracellular Tendon Matrix and Short-Term Cyclic Stretching.. Stem Cells Int 2016;2016:7342379.
- Tang JB, Wu YF, Cao Y, Chen CH, Zhou YL, Avanessian B, Shimada M, Wang XT, Liu PY. Basic FGF or VEGF gene therapy corrects insufficiency in the intrinsic healing capacity of tendons.. Sci Rep 2016 Feb 11;6:20643.
- Yang G, Rothrauff BB, Tuan RS. Tendon and ligament regeneration and repair: clinical relevance and developmental paradigm.. Birth Defects Res C Embryo Today 2013 Sep;99(3):203-222.
- Rich T, Henderson LB, Becker DL, Cornell H, Patterson-Kane JC. Indicators of replicative damage in equine tendon fibroblast monolayers.. BMC Vet Res 2013 Sep 11;9:180.
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