Development Of Cell Co-culture And Biological Microenvironment Imitation Based On Microfluidic Platform

Recently microfluidic chips have been widely used in cell research,for a lot of advantages over conventional methods like low sample and reagent consumption,versatile structures and functions,high integration,similar size to cells and in vivo microenvironment.Tremendous advances have been achieved through combining microfluidic technology with different analysis methods and integrating various structures and functions.Compared to conventional methods,the primary advantage of microfluidic chips is high integration,enabling the combination of different cells or tissues into an ordered system,which is also an important trend in current cell related research.By co-culturing different cells that are related for in vivo situation,the functions and biological characters could be better retained,which is of great significance for the reconstruction of in vitro biological models.In this thesis,some works around cell co-culture and biological environment imitation were carried out.First,the development of microfluidic technology and the applications in cell research were summarized.Specially works about cell co-culture and biological environment imitation were emphatically introduced and discussed,showing the advantages and significance of cell co-culture research on microfluidic chips.Second,an in vitro liver-tumor model was established in the microfluidic chip by co-culturing liver cancer cells and breast cancer cells.The prodrug capecitabine metabolism was successfully realized and the intermediate metabolite was successfully detected by mass spectrometry.Using this platform not only the cell vitality and proliferation could be real time monitored,the signal molecules or intermediate product could also be detected and identified by MS,showing a potential to replace conventional drug screening methods.Third,a novel approach for precisely controlled microscale cell patterning and co-culture in microfluidic chip by inkjet printing was successfully developed.Liver cancer cells and glioma cells were successfully co-patterned in the microchip and realized drug metabolism and diffusion experiment.The developed approach will help signi?cantly increase cell patterning efficacy in microfluidic chip as well as reduce theextent of laborious experimental work by automation.Fourth,a multi-functional 3D microfluidic platform was developed for co-culture of glioma cells,endothelial cells and macrophages to imitate the glioma niche at some extent.The cell morphology change,phenotype change and migration in the microenvironment could be simultaneously monitored.EMT change of glioma cells were observed under co-culture,which was consistent with in vivo situation.Cell migration and phenotype change of macrophages indicated by cytokines secretion change were also observed.