미세유체칩을 사용한 혈소판과 내피세포의 결합 및 유리 현상 관찰

Abstract

Polydimethylsiloxane (PDMS) microfluidic chip is widely adopted to simulate and model biological phenomena. We fabricated a single straight channeled PDMS microfluidic chip to replace parallel plate flow chamber in observing platelet binding to endothelial surface mediated by ultra-large von Willebrand factor (ULVWF). We cultured endothelial cells in the microfluidic channel and through which perfused buffered platelet suspension. By stimulating the endothelial cells, beads on string structures composed of platelets and ULVWF were observed on endothelial surfaces immediately after perfusing platelet suspension. The appearances were exactly the same as reported by other researchers before with parallel plate flow chambers. The number of strings observed per chip was 182.9± 59.9 (mean± SD, 102-305) from 11 chips. The blood volume required for the same results with microfluidic chips was almost a twentieth of that from parallel plate flow chambers. The workloads and time required for procedure were also considerably reduced by replacing flow chambers with microfluidic chips. The beads on string structures on the endothelial surface were promptly cleaved and washed away by perfusing buffer with added plasma. The metalloprotease activity of ADAMTS13 in the plasma was demonstrated by adding EDTA to the plasma which inhibited string cleavage. The microfluidic chip introduced here can substantially facilitate the experiments focusing on the interaction between platelets and endothelial cells.