Human EC-SMC Co-culture Based On The Microfluidics

Cardiovascular diseases including Atherosclerosis (atherosclerosis,AS) have the highest morbidity of the world, and the imbalance of vascular endotheial cells(EC) and smooth muscle cells(SMC) interaction is the important cause of these diseases. For better understanding of the pathogenesis and treatment of vascular diseases, it becomes researching hotspots to establish an appropriate model of vascular cell interaction. Currently, most of the co-culture model just mixed these two kinds of cell, and it is difficult to observe the precise mechanism of action between EC and SMC since cells easily disturb with each other; It is also hard to simulate the microenvironment or detect the single cell/sub-cellular change in living cell dynamically.Therefore, we proposed an single cell EC-SMC co-culture model based on microfluidics.By enzymatic hydrolysis and attatched tissue method from fresh umbilical cord, the vascular EC and SMC were obtained and identified by immunohistochemistry with specific antibody. Based on fluid mechanics, a "U" type of trap for cell capture was used to design the interior channel structure and cell culture chambers of the microfludics with two kinds of cell interaction models, direct contact and non-direct contact, for simulating the structure between EC and SMC in blood vessels. Ploydimethylsiloxane (PDMS) was chose as the material to make the chips with molding method. EC and SMC were processed the capture and culture experiment on two chips with different cell concentration and flow speed to observe the cell growth state under microscope.It was showed from the result that the purity of EC and SMC were both more than 99%, and with the cell concentration 3 X 105 cells/mL and the flow speed 50 μL/h, single cell could be captured on the microfluidic to build two different co-culture models.These results indicated that we successfully established the single cell EC-SMC co-culture microfludics, which provided a powerful tool for the study of different cells interaction in diseases such as AS.