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ARTIFICIAL EPITHELIAM RECONSTRUCRED BY RAFT CULTURE AND BASEMENT MEMBRANE COMPONENT AND METHOD FOR RECONSTRUCTING THEREOF
ARTIFICIAL EPITHELIAM RECONSTRUCRED BY RAFT CULTURE AND BASEMENT MEMBRANE COMPONENT AND METHOD FOR RECONSTRUCTING THEREOF
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机译：筏文化和基底膜成分重建的人工上皮细胞及其重建方法
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摘要
In order to achieve the object, the present invention, when the 80-90% growth of the fibrous cells is treated with mitomycin C to divide the culture plate to produce a feeder layer, the epithelium after separating the epithelium and connective tissue with tweezers Primary culture of epithelial cells consisting of centrifugation with 0.1% trypsin / EDTA for 15 minutes, collecting epithelial cells, dispensing into feeder layers and subcultured until medium reaches 70-80% density. Making; Neutralizing type 1 collagen with 2.2% NaHCO 3 , 0.05N NaOH and 200mmol / 1 HEPES as a neutralizing solution, mixing with fibrous cells, and incubating to induce polymerization. Culturing in a medium for 7-10 days to form a monolayer, removing the culture medium above the epithelial cells, and maintaining the outer culture of the ring at the height of the dermis to expose the epithelial cells to air for 7-10 days. Performing histopic raft culture consisting of; And providing a basement membrane component by applying the lamini and type 4 collagen on the gel, respectively, and polymerizing in a 30 ° C. incubator until polymerization to provide a basal membrane component and a three-dimensional regenerated artificial epithelial tissue. do.
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