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After recovery, please store the organoid in its maintenance medium under the correct incubation condition and medium changing process.
Protocol Diagram of liver organoid differentiation.
H&E Staining of liver organoid.
The liver organoids show expression of hepatocytes (HNF4a), hepatobiliary tracts (CK19), Albumin (ALB) and vascular endothelial cell (CD31).
Pre-mature liver organoids show liver lobules-like structure.
Liver organoids show better performance than HepG2 cell line in terms of the expression of Albumin, CYP3A4, CYP1A2, CYP2C9, CYP7A1 and BSEP.
Liver organoid show response to Rifampicin treatment as indication by the increase fluorescence intensity of CYP3A4.
At 20 days of age, the albumin secretion of ACRO's liver organoids was consistent with the secretion amount reported in the literature, being 15 pg Albumin / ng RNA. As the organoids continued to mature and grow, their albumin secretion would keep increasing, reaching a level slightly lower than that of the primary tissue.
ACRO utilized liver organoids to construct the NASH model. As can be seen from the oil red staining results shown in the above figure, the modeling group of NASH exhibited obvious lipid accumulation (red positive staining).
During the process of constructing the NASH model, we detected the expression of inflammatory factors in liver organoids through qPCR experiments, and observed the upregulation of factors such as IL-6 and IL-33. This confirmed the inflammatory response of our liver organoids.
Liver organoids can be used to detect hepatotoxicity. After treating liver organoids with the commonly hepatotoxic positive drug Trogalitazone, changes in cell viability within the liver organoids can be observed through the results of AO/PI staining.
Fluorescent-free 5-Carboxy-2',7'-Dichlorofluorescein with organoesterase activity are metabolized into fluorescent-emitting CDF, and CDF is actively transported from hepatocytes to the bile duct network and deposited in the bile duct network to simulate the cholestasis process. Trogalitazone treated organoids shows the accumulation of the CDF.
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