在 vitro 重建和有从一个 untransfected 人的 seeder 房间的设计织物的人的角膜的上皮的描述角膜的上皮的房间线

AIM:To demonstrate the morphology and structure of in vitro reconstructed tissue-engineered human corneal epithelium(TE-HCEP) with seeder cells from an untransfected HCEP cell line.· METHODS:The TE-HCEPs were reconstructed in vitro with seeder cells from an untransfected HCEP cell line,and scaffold carriers of denuded amniotic membrane(dAM) in air-liquid interface culture for 3,5,7 and 9 days,respectively.The specimens were examined with hematoxylin-eosin(HE) staining of paraffin-section,immunocytochemical staining,scanning and transmission electron microscopy.· RESULTS:During in vitro reconstruction of TE-HCEP,HCEP cells formed a 3-4,6-7 and 8-10 layers of an HCEP-like structure on dAMs in air-liquid interface culture for 3,5 and 7 days,respectively.But the cells deceased to 5-6 layers and the structure of straified epithelium became loose at day 9.And the cells maintained positive expression of marker proteins(keratin 3 and keratin 12),cell-junction proteins(zonula occludens-1,E-cadherin,connexin 43 and integrin β1) and membrane transport protein of Na+-K+ ATPase.The HCEP cells in TE-HCEP were rich in microvilli on apical surface and established numerous cell-cell and cell-dAM junctions at day 5.· CONCLUSION:The morphology and structure of the reconstructed TE-HCEP were similar to those of HCEP in vivo.The HCEP cells in the reconstructed TE-HCEP maintained the properties of HCEP cells,including abilities of forming intercellular and cell-extracellular matrix junctions and abilities of performing membrane transportation.The untransfected HCEP cells and dAMs could promisingly be used in reconstruction HCEP equivalent for clinical corneal epithelium transplantation.