title: Chondrogenic differentiation of human mesenchymal stem cells on fish scale collagen.
abstract: Fish collagen has recently been reported to be a novel biomaterial for cell and tissue culture as an alternative to conventional mammalian collagens such as bovine and porcine collagens. Fish collagen could overcome the risk of zoonosis, such as from bovine spongiform encephalopathy. Among fish collagens, tilapia collagen, the denaturing temperature of which is near 37°C, is appropriate for cell and tissue culture. In this study, we investigated chondrogenic differentiation of human mesenchymal stem cells (hMSCs) cultured on tilapia scale collagen fibrils compared with porcine collagen and non-coated dishes. The collagen fibrils were observed using a scanning electronic microscope. Safranin O staining, glycosaminoglycans (GAG) expression, and real-time PCR were examined to evaluate chondrogenesis of hMSCs on each type of collagen fibril. The results showed that hMSCs cultured on tilapia scale collagen showed stronger Safranin O staining and higher GAG expression at day 6. Results of real-time PCR indicated that hMSCs cultured on tilapia collagen showed earlier SOX9 expression on day 4 and higher AGGRECAN and COLLAGEN II expression on day 6 compared with on porcine collagen and non-coated dishes. Furthermore, low mRNA levels of bone gamma-carboxyglutamate, a specific marker of osteogenesis, showed that tilapia collagen fibrils specifically enhanced chondrogenic differentiation of hMSCs in chondrogenic medium, as well as porcine collagen. Accordingly, tilapia scale collagen may provide an appropriate collagen source for hMSC chondrogenesis in vitro.
authors.affiliation: Department of Life Science and Biotechnology, Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, 1-1-1 Higashi, Tsukuba Central-5, Ibaraki 305-8562, Japan.
authors.forename: Han-Hsiu
authors.initials: HH
authors.lastname: Hsu
authors.affiliation: Department of Life Science and Biotechnology, Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, 1-1-1 Higashi, Tsukuba Central-5, Ibaraki 305-8562, Japan. Electronic address: uemura@prec.eng.osaka-u.ac.jp.
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Uemura
authors.affiliation: Taki Chemical Co. Ltd., 346 Miyanishi, Harima-cho, Kako-gun, Hyogo 675-0145, Japan.
authors.forename: Isamu
authors.initials: I
authors.lastname: Yamaguchi
authors.affiliation: Department of Metallurgy and Ceramics Science, Tokyo Institute of Technology, S7-6, 2-12-1 Ookayama, Meguro-ku, Tokyo 152-8550, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Ikoma
authors.affiliation: Department of Metallurgy and Ceramics Science, Tokyo Institute of Technology, S7-6, 2-12-1 Ookayama, Meguro-ku, Tokyo 152-8550, Japan.
authors.forename: Junzo
authors.initials: J
authors.lastname: Tanaka
mesh_terms: D015055:1-Carboxyglutamic Acid / Q000235:genetics; D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D020219:Chondrogenesis / Q000235:genetics; D003094:Collagen / Q000378:metabolism* / Q000648:ultrastructure; D024043:Collagen Type II / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000378:metabolism; D013552:Swine; D017210:Tilapia / Q000033:anatomy & histology
chemical_list: C508319:ACAN protein, human; D053674:Aggrecans; D024043:Collagen Type II; D006025:Glycosaminoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D015055:1-Carboxyglutamic Acid; D003094:Collagen
keywords: Chondrogenesis; Extra-cellular matrix; Fish scale collagen; Mesenchymal stem cells; Tilapia collagen