Tenascin-C Expression in Equine Tendon-derived Cells During Proliferation and Migration.
In vitro cell studies might be a useful tool for studying tendon pathology, but no suitable in vitro models exist for tendon disorders. The purpose of this study was to confirm whether cell scratch culture using tendon-derived fibroblasts can provide a suitable in vitro tendon disorder model. Extracellular matrix components were examined immunohistochemically in tendon tissue, and then their related gene expression levels were analyzed by conventional reverse transcription polymerase chain reaction (RT-PCR) and/or quantitative real-time RT-PCR in tissues and cells. Collagen type I (Col I), collagen type III (Col III), tenascin-C (TN-C) and cartilage oligomeric matrix protein (COMP) were detected in tendon tissue sections, and RT-PCR confirmed their expression in tendon tissue and cells. Cells that had been cultured from explanted tendon tissue maintained the characteristics of in vivo tendon cells. The combination of TN-C and COMP might be a useful marker of tendon cells because they display more tendon-specific expression than Col I and III. In particular, the significant increase of TN-C mRNA expression in the scratch wound assay, at 12 hr after scratching, concomitant with the regeneration of the cell sheet, indicates its crucial role in tendon cell proliferation and migration. Thus, TN-C appears to be a key factor in tendon wound healing. In vitro cell scratch assays using tendon cells appear to mimic the repair of tendon tissue after injury.
Laboratory of Veterinary Physiology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan ; Present address: Epizootic Research Center, Equine Research Institute, Japan Racing Association, Shiba 1400-4, Shimotsuke, Tochigi 329-0412, Japan.
Manabu
M
Nemoto
Laboratory of Veterinary Physiology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
Keiichiro
K
Kizaki
Laboratory of Veterinary Biochemistry and Cell Biology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
Yoshio
Y
Yamamoto
Laboratory of Veterinary Physiology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan ; Present address: Department of Biological Resources, Integrated Center for Sciences, Ehime University, Shitsukawa, Toon, Ehime 791-0295, Japan.
Toshina
T
Oonuma
Laboratory of Veterinary Physiology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
Kazuyoshi
K
Hashizume
cartilage oligomeric matrix protein; collagen; equine tendon; scratch wound assay; tenascin-C