Development Of Hepatoma Piezoelectric Cell-based Sensor Chip And Application For Drug Screening

By acquiring and analying the response information from cells, the piezoelectric cell-based sensor chip (PCSC) was used to study cells or special materials. Plentiful information including growth and metabolism of cells, as well as responses to effects of complex and toxic substance can be obtained synchronously by the technology. Because of advantages in detection strategies such as being label-free, real-time, in-situ and non-invasive, this new analytical tecnology is most promisingly applicable for screening drugs, especially anti-cancer drugs.The aim of the thesis is to construct the PCSC and find the response mechanic model of cell behavior in PSCS based on theory and experiments. By the technology the application in screening drugs were explored. Not only the task had promoted the analysis methods, but had it provided potential experimental proves for clinic medicine.The cell culture technology and a piezoelectric biosensor chip were combined for the establishment of the piezoelectric cell-based sensor chip. PCSC studied HepG2 cells behaviors in real time. In order to verify the dynamic process of cells adhesion behavior, SEM and DTA assay were operated. Simultaneously MTT assay were examined to note the similarities or differences of results by PCSC and MTT.Some progresses have been achieved as follows:①Some factors that influence the stability of this analytical system were detected by PCSC , such as temperature, repeat, sensitivity. Some results were gained which were the system steability of≤0.32±0.13 Hz·min^-1 in gas phase and≤0.29±0.09 Hz·min^-1 in liquid phase, the repeat of RSD=6.55×10^-7 and the sensitivity of 340 cells.②PCSC was applied successfully in the real-time monitoring of human HepG2 cell behaviors in situ at the first time and the classic response curve was gained, which was separated 3 phases, respectively the free phase, attachment phase and steady phase according to the property of the frequency shift. The adhesion of HepG2 cells on the gold crystal surface was observed by SEM. Photos were obtained in different cell concenteations. In order to verify the corresponding connection of cells behaviors and△f , DTA was operated in PCSC and the kinetics of the trypnization was analyzed. The response highest concenteation was 106·mL-1 by analysis and calculation.③There was a line relationship between the ?f values and cell concentrations attached on the surface of PCSC. Within the response scope, the equation is...