Huh-7细胞株3D支架培养的药物代谢基因表达及对乙酰氨基酚药物毒性研究

Development of ex vivo cell culture models that adequately recreate in vivo conditions is useful in improved drug screening. Acetaminophen (APAP), well-known as paracetamol. Overdoses of APAP can induce hepatotoxicity which in turn results severe liver injury. There is a constant demand for in vitro high-throughput hepatic experimental platforms for measuring the hepatotoxicity. The purpose of this work is to provide a simple, easy-to-fabricate 3D hexagonal, hexa-channel micromachined scaffolds for studying liver cell line (Huh-7) and to provide a 3D scaffold culture system as an in vitro model for APAP induced hepato-toxicity studies on Huh-7cell line. A silicon mould based template was used to cast out polydimethylsiloxane (PDMS) hexagonal scaffolds. Cells are cultured in the scaffolds which are placed inside a well plate. RNA isolated on day 3 and day 7 of culture, gene expression on these two different time points were analyzed by qPCR. Cytoskeletal assessment is done by rhodamine-phalloidin staining and cytotoxicity was performed by MTT assay. The difference in the mRNA levels of cytochrome P450 2E1 (CYP2E1) expressions was demonstrated through qRT-PCR technique. This work shows the effects of using Polydimethylsiloxane (PDMS) hexagonal scaffolds of 50um height as a three-dimensional model to study Huh-7.