The In Vitro Study Of Tissue Engineered Artificial Esophagus

Objective:With natural tubular ECM as a scaffold and BMSCs as seed cells,BMSCs are seeded on ECM for composite culturing to investigate invitro culture of tissue-engineered artificial esophagus, thus providingexperimental basis for tissue-engineered artificial esophagus.Methods:Tubular ECM would be produced with a special scraper through aseries of processing of physical scraping and chemical methods. Themultilayer tubular ECM should be nested and modeled by using balloonand mold, creating a natural tubular ECM scaffold, and observed byscanning electron microscopy and HE staining. Irradiation sterilization istaken on the ECM scaffold by using particle accelerator; canine BMSCsare separated, extracted, cultured in vitro amplified and inducted, andalso identified with flow cytometry. And the growth curve of BMSCs isdrawn. BMSCs are seeded on ECM for composite culturing andcomparative studies are carried between BMSCs seeded on ECM andBMSCs seeded on artificial materials ePTFE. Also HE staining isobserved and compared.Results：1、Extracellular matrix is obtained after physical and chemicaltreatment, and it is a translucent white membrane with good elasticity, the natural tubular ECM is obtained by shaping with the balloon and stent,we can see the ECM has a stereo three-dimensional space structure, andthe surface without residual cell components observed by scanningelectron microscopy and HE staining. The sterilization of particleaccelerator with25KGy irradiation dose can achieve ideal effect.2、After a series of processing, centrifugal, cultured in vitro, a lot ofcell colonies separate from the canine bone marrow blood, the thirdgeneration cell were detected by flow cytometry, the result showed that:the expression rate of CD29, CD44, CD90and CD105is95.7%、98.4%、99.0%and100%respectively, while CD34, CD45almost not expressed,antigen expression in line with the BMSCs.3、BMSCs are seeded on ECM scaffold for composite culturing andobserved by HE staining, showed a large number of blue-stained nucleusstructure, each other connected closely, cell growth can also be found inthe pores. For the growth rate with BMSCs, ECM is better than artificialmaterials ePTFE.Conclusion：ECM has a good three-dimensional structure, porosity uniform andinterpenetration between each other, so it is a good choice for tissueengineering scaffolds after by shaping and sterilized. Bone marrowmesenchymal stem cells are easy to separate, extract, amplificate andidentify in vitro, and they are also have multiple differentiation potential, so they are considered as the best seed cells for tissue engineeringcurrently. BMSCs are seeded on ECM for composite culturing for tissueengineered artificial esophagus is feasible.