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Research Journal of Pharmacy and Technology
Year : 2020, Volume : 13, Issue : 5
First page : ( 2399) Last page : ( 2406)
Print ISSN : 0974-3618. Online ISSN : 0974-360X.
Article DOI : 10.5958/0974-360X.2020.00431.X

Expression of Drug-metabolizing genes and Acetaminophen drug toxicity studies on 3D scaffold culture of Huh-7 cell line

Suma M S1, Jamuna K S1, Ramesh C.K.1,*, Mahmood Riaz2

1Molecular Biomedicine Laboratory PG Dept. of Studies and Research in Biotechnology, Sahyadri Science College, Kuvempu University, Shivamogga-577203, Karnataka, India

2Department of Postgraduate Studies and Research in Biotechnology and Bioinformatics, Jnana Sahyadri, Kuvempu University, Shivamogga, 577451, Karnataka, India

*Corresponding Author E-mail: ckramck@gmail.com

Online published on 16 June, 2020.

Abstract

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.

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Keywords

Perfusion-based culture, PDMS scaffolds-based cell culture, drug metabolism gene expression, Huh-7 cell line, APAP, cytotoxicity.

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