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Kiwamu Imagawa, Kenichi Maeda, Shuichi Yokoyama, Yuki Hosoda, Shunsuke Watanabe, Takahiro Nakamura, Shigeru Kinoshita; Corneal epithelial cell bank; newly developed sheet manufacturing technology that allows the production of stable human corneal epithelial cell sheet. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5515.
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The purpose of this study is to develop cell expansion and a cryopreservation technology of corneal epithelial cells, which enables the production of stable and large number of the cell sheets from one cornea.
We cultured primary corneal epithelial cells in serum-free medium with some supplements. After detaching by enzymatic treatment, the collected cells were frozen by the addition of cryoprotectant and stored them as corneal epithelial cell bank. We conducted quality tests using post-thawed cells and produced cell sheets by culturing the epithelial cells that meet the criteria for quality tests.
It was possible to store corneal epithelial cells expanded with serum-free media in primary culture. By seeding the post-thawed cells on the amniotic membrane, we were able to produce cell sheets with stratified structure. Characteristics of the secondary cultured cell sheets, such as cell viability, electrical resistance, thickness of the cell sheet, surface antigens, were almost the same compared with the cell sheets produced by using primary cells that have not undergone the freeze-thaw process.
By developing a cryopreservation method of human corneal epithelial cells, we were able to create corneal epithelial cell bank. By increasing the number of cells in primary culture, the number of cell sheets that can be produced from the donor cornea of one eye, have increased dramatically. Cryopreservation of the cells allowed to manage a production schedules without being affected by the receiving time of the donor cornea. If we use the only cells that meet certain quality tests at cell bank, It is considered to reduce the variation in quality between the cell sheets.
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