Tracheal Epithelial Cell Cultures And Construction Of Tracheal Epithelium-free Jejunal Complex

Objective There's a difficult problem of the lung infection after the reconstruction of trachea with the jejunum. In order to solve the problem, we constructed the complex of tracheal epithelium-free jejunum, using the tracheal epithelium instead of the jejuna epithelium and cultured the tracheal epithelial cells in vitro.Methods 12 adult mongrel dogs were divided into group A(4 dogs), group B(4 dogs) and group C(4 dogs). In group A, we separated the tracheal mucosa from the dog's trachea and cut it into 0.5mm diameter and then put it on the inner face of the jejunum whose mucous layer had been removed, then used the silicone tube stand to support it independently. In group B, we cut the tracheal mucosa into 0.5mm diameter and then put it on the inner face of the jejunum whose mucous layer had been removed and used the Shape-memory Titanium-nickel Alloy stand to support it. In group C, we cut the tracheal mucosa into 5mm X 5mm and put it on the inner face of the jejunum whose mucous layer had been removed and then used the Shape-memory Titanium-nickel Alloy stand to support it.2 and 4weeks later, we obtained the complex, observed the epithelium's growing through the microscope. In addition, the tissue of tracheal mucosa and the tracheal epithelial cells purified by trypsin-digested were cultured in serum-free medium. After cultured for 2 days, the cells were observed by microscope daily and were subcultured 12days later.Results There is one dog died both in Group A and Group B because of adhesions and the infection of complex. On the inner surface of the complex of group C, we observed the growth of the epithelium, and the squamous epithelium metaplasia of the pseudostratified ciliated columnar epithelium. Neither in group A nor in group B, the same results could be observed. The tracheal epithelial cells primary cultured by the method of tissue explants grew well. After subcultured, the rells grew slowly, and died 3days later. The cells purified by trypsin-digested grew poorly, and died 3days later. Tracheal anastomosis, and intestinal anastomosis grew well without stenosis.Conclusion The trachea epithelium with appropriate size could survive on the inner surface of the jejunum whose mucous layer had been removed. The Shape-memory Titanium-nickel Alloy stand could be a good support. The tracheal epithelial cells cultured by the method of tissue explants could grow well.