Objective To study the construction feasibility of a biodegradable artificial esophagus by the squamous epithelial cells and the myoblast cells seeded on the small intestinal submucosa(SIS) and to investigate the growth patternand angiogenesis of the co-cultured human embryonic squamous epithelial cells and the skeletal myoblasts in vivo. Methods The squamous epithelial cells and the myoblast cells were obtained from the 20-week aborted fetus. Both of their cellswere marked by 5-BrdU in vitro.The isolated cells were then seeded on the SIS and co-cultured in vitro for 24 hours, and then the compound of the cells and the SIS was transplanted into the subcutaneous tissue of the athymismus mice. The observation on the morphology and the cytokeratin AE3 and α-actin specified immunohistochemistry of the squamous epithelial cells and the myoblastcells was performed at each of the following time points: 3 days, 1 week, 2 weeks, and 3 weeks after transplantation. Results The morphological observation indicated that the cultured cells could penetrate into the small intestinal submucosa and form several-layered cell structures, and that the compound of the cells and the SIS could have angiogenesis within 2-3 weeks. The 5-BrdU specified immunohistochemical observation suggested that the cells growing in the small intestinal submucosa scaffold might be the cells transplanted.The cytokeratin AE3 specified and α-actin specified immunohistochemical studies demonstrated that the transplanted cells could differentiate in vivo. Conclusion It is possible to fabricate the framework of a biodegradable artificial esophagus with the epithelial cells and the myoblast cells seeded on the small intestinal submucosa.
Objective To verify adhesion and growth ability of canine esophageal epithelial cells (EECs) on the poly (lactic-co-glycolic acid) (PLGA), a three-dimensional biodegradable polymer scaffold, and to reconstruct the canine esophagus by the tissue engineering. Methods Free canine EECs isolated from adult dogs by esophagoscopy were seeded onto the PLGA scaffolds precoated with collagen type Ⅳ after the first passage by the in vitro culture. Then, the composites of the cell-scaffold were respectively cultured invitro and in the abdominal cavity of the dog in vivo. After different periods, the cell-seeded scaffolds were assessed by histological HE staining, scanning electron microscopy, and immunohistochemical analysis. Results The cells displayed a cobblestone-shaped morphology that was characteristic of the epithelial cells and were stained to be positive for cytokeratin, which indicated that the cells were EECs. The canine EECs were well distributed and adhered to the PLGA scaffolds, and maintained their characteristics throughout the culture period. After the culture in vivo for 4 weeks, the cell-seeded scaffolds looked like tissues. Conclusion PLGA scaffolds precoated with collagen type Ⅳ can be suitable for adhesion and proliferation of EECs, and can be used as a suitable tissue engineering carrier of an artificial esophagus.
Objective To discuss the applycation possibility of themicroscopic stripping technique used in the primary culture of human embryonicesophagus squamous epithelial cells, and of the methodds for the isolation, depuration and subculture of the esophagus epithelial cells in vitro. Methods The squamous epithelial cells wereobtained from the esophagus mucous membrane of the 20-week abortion fetus through the microscopic stripping technique, and were digested with trypsin. Then, the morphological, immunohistochemical observation and the growth curve of the isolated cells were studied. Results The isolated cells were spherical in the cell suspension and spherical-like or polygon-like after attachment to the culture flask.The squamous epithelial specialized cytokeratin staining was bly positive. And the morphological studies by the transmission electron microscopy indicated that the cultured cells were squamous epithelial cells. The squamous epithelial cells reached the peak level 3-4 days after the transfer of the culture. The absorbanceat 3 and 4 days was significantly higher than that at 1,2,5 and 6 days (P＜0.05). Conclusion A large mumber of squamous epithelial cells can be available with the microscopic stripping technique and the digestion method. Thecultured squamous epithelial cells can be proliferated quickly, and fit for the tissue engineering study.
ObjectiveTo review the research progress of the tissue engineering technique in the esophageal defect repair and reconstruction. MethodsThe recently published clinical and experimental literature at home and abroad on the scaffold materials and the seeding cells used in the tissue engineered esophageal reconstruction was consulted and summarized. ResultsA large number of basic researches and clinical applications show that the effect of the tissue engineered esophagus is close to the autologous structure and function of the esophagus and it could be used for the repair of the esophageal defect. However, those techniques have a long distance from the clinical application and need an acknowledged rule of technology. ConclusionTissue engineering technique could provide an innovative theory for the esophageal defect reconstruction, but its clinical application need further research.