Premium
Differentiation of HepG2 and Huh7 cells using dimethyl sulfoxide
Author(s) -
Mowbray Catherine Anne,
Howard Alison,
Morsman Janine,
Hirst Barry H.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb654
Subject(s) - dimethyl sulfoxide , hepatocyte , cell culture , messenger rna , albumin , microbiology and biotechnology , cyp3a4 , biology , cell , chemistry , andrology , enzyme , in vitro , biochemistry , cytochrome p450 , gene , medicine , genetics , organic chemistry
The secondary hepatocytes HepG2 and Huh7 are phenotypically poorly differentiated compared with hepatocytes in situ , limiting their use in drug development. This study assessed the efficacy of dimethyl sulfoxide (DMSO) to promote mature hepatocyte‐like differentiation. mRNA of two hepatocyte maturity markers (albumin (ALB) and transferrin (TF)), one of immaturity (α‐fetoprotein (AFP)), and key metabolic enzymes CYP3A4 and UGT1A1, was measured by qPCR. Responses to DMSO were normalised to confluent untreated cells (CUC). 1% DMSO increased ALB and TF mRNA in both cell lines with maximal expression at 15 to 20 days, followed by a return to control levels (eg. HepG2: ALB: confluent untreated cells (CUC): 1.0 ± 0.1; 15d: 6.6 ± 2.3, p<0.05. TF: CUC: 1.0 ± 0.2; 20d: 13.8 ± 0.6, p<0.05). DMSO did not reduce AFP mRNA in either cell line. CYP3A4 mRNA was low in both cell lines and not affected by DMSO. 2% DMSO increased UGT1A1 mRNA in both cell lines (HepG2: CUC: 1.0 ± 0.2; 5d: 2187.0 ± 153.0, p<0.05. Huh7: CUC: 1.0 ± 0.2; 28d: 4.0 ± 0.3, p<0.05). The data indicate these cell lines may effectively be differentiated to provide suitable models for early stage drug testing. Funded by BBSRC and Sanofi‐Aventis.