GenJet™ In Vitro DNA Transfection Reagent for HepG2 Cells

Description
GenJet™ DNA In Vitro Transfection Reagent for HepG2 Cells is specifically optimized for efficient plasmid DNA delivery into HepG2 cells.

HepG2 is a well-characterized human hepatocellular carcinoma cell line derived from the liver tissue of a 15-year-old male donor with a differentiated hepatocellular carcinoma. The cells exhibit epithelial morphology and retain many liver-specific functions, making them one of the most widely used in vitro models for studies of hepatic metabolism, gene regulation, drug discovery, toxicology, viral infection, and recombinant protein expression. HepG2 cells synthesize and secrete numerous plasma proteins, including albumin, transferrin, α1-antitrypsin, α2-macroglobulin, and plasminogen, and respond to a variety of hormonal and metabolic stimuli. Their robust growth characteristics and adaptability to large-scale culture systems have further contributed to their extensive use in biomedical and pharmaceutical research.

The transfection conditions described below have been optimized to achieve high transfection efficiency in HepG2 cells while maintaining excellent cell viability. One milliliter of GenJet™ Reagent is sufficient for approximately 300–600 transfections in 24-well plates or 50–100 transfections in 6-well plates, depending on the culture format and amount of DNA used.

Storage Conditions
Store at 4°C. When maintained under the recommended storage conditions, the product remains stable for at least 24 months.

Representative Transfection Data in HepG2 Cells

GenJet™ Reagent is specifically optimized for HepG2 cells (ATCC® HB-8065™) and demonstrates superior transfection performance compared with Lipofectamine™ 2000 (L2K) and the Amaxa® electroporation system. HepG2 cells were cultured according to ATCC-recommended conditions on collagen type I–coated culture dishes and transfected with the GFP reporter plasmid pEGFP-N3. Transfection efficiency was evaluated 48 hours post-transfection.

Right Panel: Comparison of transfection efficiencies achieved with GenJet™, Lipofectamine™ 2000 (L2K), and the Amaxa® electroporation system in HepG2 cells. A 4.7-kb GFP expression plasmid (pEGFP-N3) was delivered into HepG2 cells cultured on collagen-coated dishes according to each manufacturer’s recommended protocol. Transfection efficiency, expressed as the percentage of GFP-positive cells, and GFP fluorescence intensity were quantified by flow cytometry (FACS) 48 hours post-transfection.

Left Panel: Effect of serum and antibiotics on GenJet™-mediated transfection in HepG2 cells. Cells cultured on collagen-coated dishes were transfected under three different conditions: (1) serum- and antibiotic-free medium; (2) medium containing 10% serum and antibiotics, followed by medium replacement 5 hours post-transfection; and (3) medium containing 10% serum and antibiotics without medium replacement following transfection. The presence of serum and antibiotics was found to enhance GenJet™-mediated gene delivery and transgene expression in HepG2 cells.

Representative images demonstrating the transfection performance of the pre-optimized GenJet™ DNA Transfection Reagent in HepG2 cells (ATCC® HB-8065™). HepG2 cells were cultured under ATCC-recommended conditions on collagen type I–coated culture dishes and transfected with either the GFP reporter plasmid pEGFP-N3 (4.7 kb; right panel) or the β-galactosidase expression plasmid pSV-β-galactosidase (6.9 kb; left panel). Transgene expression was evaluated 48 hours post-transfection by confocal fluorescence microscopy using a Zeiss LSM 510 system for GFP detection and by β-galactosidase staining for assessment of β-galactosidase expression. The results demonstrate efficient delivery and expression of both reporter genes in HepG2 cells.

Transfection Protocol