Figure 7 Cell viability and morphology in liver-on-a-chip. (A) Schematic of cell encapsulation and seeding. (B-D) Representative images showing living cells (green: Calcein AM) and dead cells (red: PI) at Day 4. Scale bar=200 µm. (E) Cell viability at Day 0 to Day 4. (F) Comparison of burst pressure of the chips made by layer-by-layer adhesion and sacrificial template. (G) Maximum shear stress that the chips were able to endure.
In previous study, poly-L-lysine has been used as an electrostatic glue to seal the hydrogel-based chips 32. However, this method provided weak adhesion with fluid leakage occurring at applied pressures higher than 3 kPa, providing an extremely low shear stress of approximately 0.1 dyne/cm2. To compare the perfusion tolerance of chips constructed by different methods, liver-on-a-chip with the same channel morphology was constructed by Gel-MA hydrogel and alginate sacrificial template. As shown in Figure 7F, the chip made by layer-by-layer adhesion showed the high burst pressure of >40 kPa during the culture, due to the strong support provided by the tough F127-DA layer 33. In contrast, the Gel-MA chip made by sacrificial template was brittle and weak, and was easily broken by an extremely low pressure of 3 kPa. Consistently, the assembled chip shown in Figure 7A was able to withstand flow-induced shear stress of >45 dyne/cm2, which is in the range of physiological values of 1-50 dyne/cm2 in human blood vessels 34. However, the Gel-MA chip made by sacrificial template could only tolerate a very low shear stress of 0.1-0.2 dyne/cm2, thus limiting its applications in perfusing culture. As the accurate mimicking of mechanical microenvironments is essential in construction of organs-on-chips35, layer-by-layer adhesion performs better than the use of sacrificial templates.