In vitro tissue engineering of lamellar cornea using human amniotic epithelial cells and rabbit cornea stroma

oleh: Xiao-Yong Liu, Jian Chen, Qing Zhou, Jing Wu, Xiao-Ling Zhang, Li Wang, Xiao-Yan Qin

Format: Article
Diterbitkan: Press of International Journal of Ophthalmology (IJO PRESS) 2013-08-01

Deskripsi

<b>AIM:</b>To reconstruct the lamellar cornea using human amniotic epithelial (HAE) cells and rabbit cornea stroma <i>in vitro</i> using tissue engineering technology.<b>METHODS:</b> 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 2 weeks. 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 keratin 3 and keratin 12 were detected with immunofluorescence microscopy.<b>RESULTS:</b>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.<b>CONCLUSION:</b> Functional tissue-engineered lamellar corneal grafts can be constructed <i>in vitro</i> using HAE cells and rabbit corneal stroma.