In the ICG uptake assay, bioprinted structures or cells were cultured for 48 hours in a medium supplemented with 8-bromo-cAMP, pregnenolone-16α-carbonitrile, and progesterone, as per the manufacturer's instructions. Subsequently, they were coincubated with ICG (1 mg/ml) at 37°C for 1 hour, washed three times with PBS, and then switched to HEM culture medium. The uptake/release of ICG was observed under a microscope. Bioprinted structures or cells were stained with ac-LDL (Yeasen) and periodic acid-Schiff (Solarbio), and the staining results were observed under a microscope. Different inducers were used to treat 3DP-liver and eHep cells for 48 hours, and the gene expression levels of Cyp1a2 (3-methylcholanthracene, 10 μM, Sigma-Aldrich), Cyp3a11 (rifampicin, 25 μM, Solarbio), Cyp2e1 (rifampicin, 25 μM, Solarbio and ethyl alcohol; 0.5%, v/v), Cyp3a41b (3-methylcholanthracene, 10 μM, Sigma-Aldrich), and Cyp1a1 (rifampicin, 25 μM, Solarbio and barbituric acid, 10 μM, Aladdin) were detected. The P450-Glo Cyp1a2 Induction/Inhibition Assay (Promega) was used, following the manufacturer's instructions, to detect Cyp1a2 enzyme activity in different states.
Deng B., Ma Y., Huang J., He R., Luo M., Mao L., Zhang E., Zhao Y., Wang X., Wang Q., Pang M., Mao Y., Yang H., Liu L, & Huang P. (2024). Revitalizing liver function in mice with liver failure through transplantation of 3D-bioprinted liver with expanded primary hepatocytes. Science advances, 10(23).
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