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.
OBJECTIVE
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.
Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61802, USA.
Sushmitha S
SS
Durgam
Allison A
AA
Stewart
Holly C
HC
Pondenis
Angela C
AC
Yates
Richard B
RB
Evans
Matthew C
MC
Stewart
D000818:Animals; D015152:Blotting, Northern / Q000662:veterinary; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods* / Q000662:veterinary; D024042:Collagen Type I / Q000096:biosynthesis; D024061:Collagen Type III / Q000096:biosynthesis; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000096:biosynthesis; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006736:Horses / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D064235:Matrilin Proteins; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D013710:Tendons / Q000166:cytology* / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism
D024042:Collagen Type I; D024061:Collagen Type III; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D006025:Glycosaminoglycans; D064235:Matrilin Proteins; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D007334:Insulin-Like Growth Factor I