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K. Nishitsuka, Y. Kashiwagi, T. Yamamoto, H. Yamashita; Cloning and Characterization of a Cell Strain Derived From Human Hyalocytes. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2705.
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In this study, we have established the cell strains derived from human vitreous (human hyalocytes) and compared their biological feathers with the porcine vitreous-derived cell line (porcine hyalocytes).
This research was approved by the Ethical Committee, Yamagata University Faculty of Medicine. Human vitreous specimens were obtained from two patients after securing the written permission, and cultured with 10%FBS/DMEM. To immortalize the cells, human papilloma virus (HPV) 16 E6 and E7 were transfected. To characterlize and compare the human hyalocytes and the porcine hyalocytes, we examined expression pattern of several genes by RT-PCR. To investigate the regulation of hyaluronan production, expression of hyaluronan syntase (HAS) isoforms by RT-PCR. In addition, hyaluronan production was measured after the stimulation of TGF-β1 and/or PDGF-BB using ELISA.
We obtained two types of cell strains derived from human vitreous tissue, HH64 and HH65. We isolated 4 clones of HH64 and 7 clones of HH65. The morphological features of HH64 cells resembled endothelial cells, and those of HH65 cells fibroblastic cells. The doubling time of HH64 cell strain was 58 hours and that of HH65 was 80 hours. In all the cell strains, collagen type I alpha 1 (COL1A1), collagen type II alpha 1 (COL2A1), CD11b, CD14, CD68, CD204 and CD206 were expressed at mRNA levels, and glial fibrillary acidic protein (GFAP), S-100 protein beta chain (S-100b), or CD163 were not expressed at mRNA levels. Stimulation with TGF-β1 and/or PDGF-BB did not induce hyaluronan synthase 2 (HAS2) at mRNA level or HA production. In the porcine hyalocytes, GFAP and HAS2 were expressed at mRNA levels, and HA production was inducedby TGF-β1 and/or PDGF-BB.
The human hyalocyte cell strains have macrophage-like and histiocyte characteristics, and are different from the porcine hyalocyte cells.The results suggest that variety of origins of hyalocytes in different animal species.
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