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C Yuan, Y Sawada, CS Boehlke, G Adams, AJ W Huang; Growth of Epidermal Keratinocytes on Human Amniotic Membrane . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1682.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:Ex vivo expansion of autologous limbal epithelium containing presumed corneal epithelial stem cells, has been used successfully for ocular surface reconstruction. It can circumvent the potential risks of tissue rejection in allografts. However, corneal epithelial stem cells have yet to be identified and the availability of autologous tissue is limited. We further investigated the feasibility of culturing human epidermal keratinocytes on preserved amniotic membrane to serve as stem cell alternatives for ocular surface reconstruction. Method: Discarded skin tissues from head and neck surgeries were treated with dispase. The dissociated keratinocytes were cultivated in a serum-free medium for propagation. Preserved amniotic membranes were pretreated with dispase to remove the amniotic epithelia and coated with fibronectin before use. Epidermal keratinocytes and human limbal epithelium (corneal keratinocytes) were seeded on the amniotic membranes. The growth of keratinocytes was quantified and a battery of monoclonal anti-keratin antibodies was also used to examine the differentiation of keratinocytes. Results:Both human epidermal and corneal keratinocytes attached and propagated well on the dispase-treated amniotic membranes. More interestingly, precoating with fibronectin further facilitated the focal attachment of keratinocytes onto the amniotic membrane. Epithelial differentiation of epidermal and corneal lineages was also confirmed with the expression of cytokeratin K10, K13, K19 and K3. Conclusion:Amniotic membrane can support the growth of human epidermal as well as corneal keratinocytes. The preserved epithelial differentiation of epidermal keratinocytes suggests that the epidermal lineages can be maintained on the amniotic membrane and it may be used as an ocular surface epithelial substitute.
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