Bladder tissue engineering includes three components, namely cell scaffold, seeding cells and message factor. Seeding cells is essential to construct tissue engineering organs and is the native material of organ reconstruction. It is important to find a seeding cell which can not only provide excellent biocompatibility, but also can generate enough cells in work of organ reconstruction. At present, there are some commonly used seeding cells, for example, urinary bladder transitional epithelial cells,smooth muscle cells, coculture of smooth muscle cells and transitional epithelial cells, mesenchymal stem cells. Among the total,coculture of smooth muscle cells and transitional epithelial cells and mesenchymal stern cells are better than the others. The current problems of seeding cells are as follows: ①The mature cells that have been completely differentiated are limited in source and immunological rejection. ② The mature cells that have been completely differentiated during the differentiated only can generate in the limited passage culture procedure and they will age and die out of the passage maximum; in addition, the dedifferentiation also exists; ③further analysis should focus on the coculture of smooth muscle cells and transitional epithelial cells and mesenchymal stem cells into normal bladder.

Objective To construct eukaryotic expressing vector of mouse soluble CD160 and stably transfect into CHO cells for eukaryotic expression.Methods Recombinant soluble CD160(rsCD160) was constructed by gene recombination.Total RNA was extracted from the spleen of C57BL/6 mice.cDNA was amplified for the soluble form of CD160.Then,the PCR product was cloned tO pcDNA3.1 and pEGFP-N1.The recombinant plasmid was identified by restriction map and sequence analy-sis.The soluble CDl 60 expression in CHO cells transfected with recombinant psCDl 60 was verified by RT-PCR and Westernblot.The binding ability of psCD160 tO its ligand was detected by FACS.Results 520 bp mouse soluble CD160 gene was obtained.Recombinant mouse psCD160 was successfully constructed.After transfection,soluble CD160 expression in the culture supernatant of CHO cells was successfully detected.FACS analysis indicated that soluble CD160 could bind tO its ligand.Conclusion Recombinant mouse psCD160 is successfully constructed,which will benefit our further study on soluble CD160 for immune therapy against tumor in the future experiments.

Cardiovascular Diseases ; epidemiology ; Humans ; Metabolomics