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International Journal of Bioprinting 3D-printed micro-perfused culture device

Figure 7. Cell viability test of Huh7.5 hepatocellular carcinoma cells on TCPS (left), static culture (center), and micro-perfused culture (MPC) device
(right). Confocal Z-stack images after 4 days of culture. The live cells were stained with calcein-AM (green) and dead cells were stained with ethidium
homodimer-1 (red).
the conventionally reported 200 µm viable thickness. Bean a thicker cell construct. Hence, pore size and porosity are
et al. reported that the pore size of scaffold increases with monitored in this study.
increasing fiber diameter, shedding some light that fiber CFD simulation, Flow3D, was performed to optimize
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diameter and pore size are interrelated. Conflicting studies
have been reported on the effect of fiber diameter on the the dimensions on the perfusion culture where the shear
differentiation of mesenchymal stem cells toward cartilage stress on the MPC device had to be below 0.2 Pa to prevent
phenotype where one group reported that a fiber diameter any detrimental effect on cells, especially primary liver
of 500 nm in diameter better promoted cartilage phenotype hepatocytes. 53,63 From the CFD simulation results shown
differentiation when compared to a fiber diameter of in Figure 4, as the channel width decreases, the shear
3 µm. Another study reported that using poly(L- stress increases significantly, which is in concordance to
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lactide) microfibers better promoted chondrogenesis the simple 2D Poiseuille flow system equation. From the
than nanofibers. However, the common findings among simulation studies, channel width of 800 µm and flowrate
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these studies are that large scaffold pore size promotes cell below 10 mL/h were found to exhibit acceptable shear stress
penetration into the scaffold. The porosity of the scaffold along the channel width and culturing chamber. However,
is also needed to ensure delivery of medium to the core of a lower perfusion rate is preferred to reduce excessive

Volume 10 Issue 1 (2024) 153 https://doi.org/10.36922/ijb.0226