In Vitro Model on Glass Surfaces for Complex Interactions between Different Types of Cells

Abstract

This report establishes an in vitro model on glass surfaces for patterning multiple types of cells to simulate cell-cell interactions in vivo. The model employs a microfluidic system and poly(ethylene glycol)-terminated oxysilane (PEG-oxysilane) to modify glass surfaces in order to resist cell adhesion. The system allows the selective confinement of different types of cells to realize complete confinement, partial confinement, and no confinement of three types of cells on glass surfaces. The model was applied to study intercellular interactions among human umbilical vein endothelial cells (HUVEC), PLA 801 C and PLA801 D cells.

Diffusion of self-assembled monolayers of thiols on the gold surfaces covered with polydimethylsiloxane stamps

“…We selected HS(CH 2 ) 11 (OCH 2 CH 2 ) 3 OH (EG 3 ) and HS(CH 2 ) 11 (OCH 2 CH 2 ) 6 OH (EG 6 ) as examples to illustrate our strategy. In general, thiol SAMs growth on gold substrate from solution includes three steps: the first step of the chemisorption of the headgroup (SH), the fastest step; the second step of the straightening of the hydrocarbon chains to form lying-down phase, the slower step; the third and final step of the reorientation of the terminal groups to form functional standing-up phase, the slowest step [22].…”

“…SAMs of standing-up phase with the terminal groups (OH) exposure can be detected with cells as the indicator in our experiments. Only well-ordered SAMs with the polyethylene moiety of the EG 3 or EG 6 exposed can effectively resist cell adhesion. We identified the area without cell adhesion covered with thiol SAMs as inert area and the area with cell adhesion covered with gold as active area.…”

“…We identified the area without cell adhesion covered with thiol SAMs as inert area and the area with cell adhesion covered with gold as active area. To measure the inert areas on surface, we introduced an aqueous solution of EG 3 or EG 6 into a channel with a width of 200 lm and allowed it to incubate at room temperature for 3, 5, 7, 9, 20, 30, and 40 min and then rinsed the channel with vast PBS solution at once to get rid of residual solution of EG 3 or EG 6 in order to avoid residual thiol further diffusing (Fig. 1a, b).…”

“…Fabrication of the master for microchannels was accomplished using established methods [6]. Patterns used in these experiments were designed by L-edit software and exported as a PS file.…”

“…In recent years, SAMs found wide applications in chemistry, physics, and especial biology [2][3][4][5] when SAMs combined with microfluidic technology [6][7][8][9][10]; for example, we applied alkanethiolate SAMs on gold surface to study the relationship between the shape of mammalian cells and their migration direction [11]. Recently, when we applied alkanethiolate SAMs combined with microfluidic technology to establish interaction models for multiple types of cells [12], we found that the SAMs diffused on the surfaces of evaporation-coated gold slides which were covered with polydimethylsiloxan (PDMS) stamps [13].…”