在薄片状的角膜的 vitro 织物工程使用人的羊膜的上皮的房间和兔子角膜基质

• 作者: Jian Chen Jing Wu Li Wang Qing Zhou Xiao-Ling Zhang Xiao-Yan Qin Xiao-Yong Liu
• 来源期刊: 国际眼科杂志：英文版
• 学科: 医学
• 关键词: amniotic epithelial cells CORNEA tissue engineering KERATIN
• 年，卷（期）: 2013,（4）
• 页码范围: 425-429
• 页数: 5页
• 分类号: R779.6

摘要

AIM:To reconstruct the lamellar cornea using human amniotic epithelial(HAE) cells and rabbit cornea stroma in vitro using tissue engineering technology.·METHODS:Human amnia taken from uncomplicated caesarean sections were digested by collagenase to obtain HAE cells,and the cells were cultured to proliferate.Rabbit corneal epithelial cells were removed by n-heptanol to make lamellar matrix sheets.The second passage of HAE cells were cultured on the corneal stroma sheets for 1 or 2 days,then transferred to an air-liquid interface environment to culture for 2weeks.Tissue engineered lamellar cornea(TELC)morphology was observed by Hematoxylin-eosin(HE)staining;its ultrastructure was observed by transmission electron microscopy(TEM) and scanning electron microscopy(SEM);corneal epithelial cell-specific keratin3 and keratin 12 were detected with immunofluorescence microscopy.·RESULTS:HAE cells grew on the rabbit corneal stroma,forming a monolayer after 1-2 days.About 4-5 layers of epithelial cells developed after 2 weeks of air-liquid interface cultivation,a result similar to normal corneal epithelium.Rabbit corneal stromal cells were significantly reduced after one week,then almost completely disappeared after 2 weeks.TEM showed desmosomes between the epithelial cells;hemidesmosomes formed between the epithelial cells and the basement membrane.SEM revealed that the HAE cells which grew on the lamellar cornea had abundant microvilli.The tissue-engineered cornea expressed keratin 3 and keratin 12,as detected by immunofluorescence assay.·CONCLUSION:Functional tissue-engineered lamellar corneal grafts can be constructed in vitro using HAE cells and rabbit corneal stroma.