Biological Application of Widefield Surface Plasmon Resonance Microscope to Study Cell/Surface Interactions and the Effect of TGF-β3, HCL and BSA/HCL on Cell Detachment Assay of Bone Cells Monolayer

摘要

Widefield Surface Plasmon Resonance (WSPR) microscope was used to investigate cell surface interactions under two different culture conditions: bone cells cultured on SPR substrate with transforming growth factor β3 (TGF-β3) and without as control. Trypsinisation was carried out in order to investigate its effect on cell detachment, in the presence of TGF-β3, HCl or BSA/HCl solutions. Trypsin was therefore added to four groups of bone cells with addition of TGF-β3, HCl, HCl/BSA solutions and one additional flask as control. These results further confirmed that application of TGF-β3, HCl and HCl/BSA decreased the degree of cell attachment on surface of culture flasks. HCl and BSA/HCl were tested as they are the carriers and solvents for TGF-β3. Cell detachment in control was about 43% after 6 min, which is slow. Bone cells in the presence of BSA/HCl started detaching from the surface about 4-5 min and cell detachment was about 63% after 6 min which was faster as compared to the control. Bone cells in the presence of HCl alone started detaching from the surface about 2 min (after applying trypsin) and cell detachment was about 69% after 6 min which was faster compared to the BSA/HCl and control. Trypsinisation experiments for bone cells cultured with TGF-β3 (50 ng/ml) showed that cells started to detach from the surface about 1 min after application of trypsin and cell detachment was about 85% after 4 min which was faster as com-pared to the control, HCl and BSA/HCl.