The objective of this study were to establish a cell line derived from porcine hyalocytes and to investigate the regulation of hyaluronan (HA) synthesis in response to cytokines. After 50 passages of the cells derived from porcine vitreous tissue, a cell line was generated. The immortalized cells showed fibroblastic morphology. The cell doubling time was 56.9 h. In the mRNA level, the cells expressed plate-derived growth factor (PDGF) α receptor, PDGF β receptor, transforming growth factor-β (TGF-β) type I receptor, TGF-β type II receptor, CD44, collagen type I, collagen type II, glial fibrillary acidic protein (GFAP), hyaluronan synthase (HAS) 2, HAS 3 and β-actin. In the protein level, GFAP was expressed in this cell line. S-100 protein and cytokeratin were not detected. Stimulation with TGF-β1 and/or PDGF-BB induced a marked increase in the expression level of HAS2 mRNA, and induced HA production. TGF-β1 stimulated HAS2 expression through the signal transduction pathway including Smad 2,3,4. In summary, this report constitutes the first successful immortalization of porcine hyalocyte cells. The production of HA was induced from the generated porcine hyalocyte cell line under the stimulation of TGF-β1 and/or PDGF-BB, which may be related to the pathogenesis of proliferative membrane formation in proliferative vitreo-retinal diseases. © 2007 Elsevier Ltd. All rights reserved.
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