Preparation Of Temperature-responsive Substrate And Its Impact On The Growth And Adhesive Behavior Of Normal Smooth Muscle Cells And Liver Tumor Cells

Organ transplantation has long been restricted by the shortage of donors and the high risks of grafted rejection. Tissue engineering which is the new tool of regenerative medicine aims to overcome the limitation of organ transplantation. However, conventional tissue engineering using biodegradable scaffolds or injecting single cell suspension has lots of disadvantanges. The biocompatibility of biodegradable scaffolds need to be test, and the degradation of scaffolds can result in inflammation. Also, the degradation will leave a lot of space in vivo which will be filled by proliferate cells and large amount of cell matrix. However, too much cell matrix and residue of scaffolds after degradation will result in tissue fiberation. As a result, researchers create new method to fabricate tissue, using temperature-responsive polymer to create temperature-responsive dishes, and then apply them in the use of tissue engineering.This paper first studied the primary culture and identification of airway smooth muscle cells. And then we used poly N-isopropylacrylamide to make temperature-responsive substrate in the basis of normal cell culture dishes, cultured airway smooth muscle cells on the top of them, and observed the impact of temperature-responsive dishes on growth and adhesive behavior of airway smooth muscle cells and liver tumor cells. At last, we try to get cell sheets by temperature-responsible dishes. Our main aim was to master the primary culture and identification of airway smooth muscle cells and the fabrication of temperature-responsive substrate, and make a theory basis for the future work.The following are the main results of this study:①We used water and isopropyl alcohol as the solvent of N-isopropylacrylamide. And we found that the water solvent can not achieve making the temperature–responsive substrate in that the concentration of N-isopropylacrylamide in water solution is too low. So that the N-isopropylacrylamide can not form covalent bond with the surface of the normal cell culture dishes.②By using theγ-irradiation of irradiation center in Southwest Hospital and the electron-beam irradiation of Sichuan research center of atom energy, we found that theγ-irradiation can only make poly (N-isopropylacrylamide) hydrogel but not the temperature-responsive substrate. The cause is that the dose rate ofγ-irradiation is too low and take up a lot of time so that the isopropyl alcohol solvent evaporate out. However, the electron beam irradiation solved this problem well.③We maked N-ispropylacrylamide in isopropyl alcohol solution with different concentration 40%,45%,50%,55%. And we found that cell adhere better on the temperature-responsive substrates made by 45% N-ispropylacrylamide isopropyl alcohol solution than other three concentration. In addition, 200μL is the best inject amount of N-ispropylacrylamide solution.④We seeded the airway smooth muscle cells and tumor cells growth on the temperature-responsive substrates and normal cell culture dishes. And we found that airway smooth muscle cells grew worse than normal cell culture dishes, however, the liver cells seeded onto the temperature-responsive substrates grew as well as on the normal cell culture dishes.⑤By using the temperature-responsive substrates to control the adhesion and detachment of the airway smooth muscle cells and liver tumor cells, we found that the airway smooth muscle cells can detach from the surface of the temperature-responsive dishes, however, the tumor cells can not manage it.