Experimental Study On Transfecting Gene Into Human Esophageal Epithelial Cells To Develop Tissue-engineered Esophagus In Vitro

Objective: To study the methods of promoting the adhesion of seeding cells to the scaffod of TEE. Methods: (1) Construction of recombinant adenovirus vector encoding FnCBD64 gene. (2)Isolation, culture and proliferation of human esophageal epithelial cells (HEECs ) and expression, secretion of the protein FnCBD₆4 after transfection of Ad.FnCBD₆4. (3)A kind of highly porous Poly (lactic-co-glycolic) acid (PLGA) scaffold was constructed by the method of particul ate leaching and then assessed by histological analysis and electronic microscopy detection. We study the effect of enhancing the adhesion of HEECs to PLGA scaffods after precoating FnCBD₆4 on scaffolds or transfecting Ad.FnCBD64 into HEECs. Results: The efficient and reliable recombinant adenovirus coding for FnCBD₆4 gene were constructed successfully. HEECs can express and secrete FnCBD₆4 efficiently after transfected with corresponding gene. The PLGA scaffold has suitable structure for growth of seeding cells. HEECs formed a continuous layer closely on the leaf-shaped PLGA scaffolds in groups of precoating FnCBD₆4 and transfecting Ad.FnCBD₆4, and both of them revealed a significant increase of cell number compare with other groups. What is more, the effect of transfecting Ad.FnCBD₆4 group is more obvious. Conclusions: HEECs harvested and proliferated in vitro can be transfected efficiently by recombinant adenovirus coding for FnCBD₆4 gene. Precoating FnCBD₆4 can promote proliferation and adhesion of HEECs on the highly porous biodegradable polymer scaffold of PLGA, and transfecting Ad.FnCBD₆4 makes better effects.