DNA transfection of primary hepatocytes
|Number of transfections|
0.5 ml of jetPEI®-Hepatocyte transfection reagent is sufficient to perform up to 150 transfections in 24-well plates or 30 transfections in 60-mm plates
5°C ± 3°C, for up to 12 months
150 mM NaCl
jetPEI®-Hepatocyte is a DNA transfection reagent designed to transfect Hepatocyte-like cells. jetPEI®-Hepatocyte is recommended to transfect primary hepatocytes and cell lines such as human hepatocarcinoma Hep G2 and murine hepatocytes BNL-CL.2.
jetPEI®-Hepatocyte is a Galactose-bearing linear polyethylenimine designed to enhance the transfection of cells expressing Galactose-specific membrane lectins, such as hepatocytes that express the asialoglycoprotein receptor (ASGP-R or Gal/GalNAc receptor). Cell targeting is the result of binding of the Galactose residues to the specific cell-surface receptors, leading to internalization of the DNA complexes.
|Reference Number||Amount of reagent||Amount of NaCl|
|102-05N||0.5 ml||50 ml|
jetPEI®-Hepatocyte leads to improved transfection efficiency of hepatocarcinoma Hep G2 cells
Compared to our standard jetPEI® transfection reagent, jetPEI®-Hepatocyte provides significantly higher transfection efficiencies in human hepatocarcinoma Hep G2 cells as well as in difficult to transfect primary human hepatocytes (Fig. 1).
Fig. 1: Comparative transfection efficiency of jetPEI®-Hepatocyte versus jetPEI®. Hep G2 (50,000 cells in 24-w) and primary human hepatocytes (100,000 cells in 24-w) were transfected using 3.2 µl of both reagents with 1 µg of pCMVEGFPLuc, in 1 ml of complete medium containing 10% of serum. The luciferase activity was determined 48h post-transfection. Transfection efficiency was expressed as relative efficiency obtained with jetPEI® compared to jetPEI®-Hepatocyte.
Good transfection efficiency and excellent viability in primary hepatocytes
Primary human hepatocytes show good transfection efficiency (around 50%) 24 h and 48 h after transfection using jetPEI®-Hepatocyte and luciferase plasmid pCMVEGFPLuc. In addition, cells look very healthy (Fig. 2).
Fig. 2: Primary human hepatocytes expressing GFP 24 h and 48 h after transfection using jetPEI®-Hepatocyte. Primary human hepatocytes (100,000 cells in 24-w) were transfected with 1 µg of pCMVEGFPLuc at N/P ratio of 8, in 1 ml of complete medium containing 10% of serum. Cells were visualized by fluorescence microscopy.
Easy transfection of hepatocyte-derived cell lines
The most popular hepatocyte-derived cell lines BNL-CL.2 and Hep G2 were successfully transfected using jetPEI®-Hepatocyte and commonly show approximately 50 % and 30% transfection efficiency, respectively (Fig. 3 and 4).
Fig. 3: BNL-CL.2 cells expressing ß-galactosidase after transfection using jetPEI®-Hepatocyte. BNL-CL.2 cells (50,000 cells in 24-w) were transfected using 3.2 µl of reagent with 1 µg of pCMV-betaGal, in 1 ml of complete medium containing 10% of serum. ß-galactosidase gene expression was visualized by X-gal staining after 24h.
Fig. 4: Hep G2 cells expressing GFP 72 h after transfection using jetPEI®-Hepatocyte. Hep G2 cells (50,000 cells in 24-w) were transfected using 3.2 µl of reagent with 1 µg of pCMVEGFPLuc, in 1 ml of complete medium containing 10% of serum. Cells expressing GFP protein were visualized by fluorescence.
Easy to use protocol
The jetPEI®-Hepatocyte protocol is as simple as the jetPEI® one: Mix the DNA with the reagent to form complexes and simply add the mixture to the cells. jetPEI®-Hepatocyte is compatible with serum and antibiotics, thus eliminating the need for media changes. Protein expression is determined 24 h to 72 h post-transfection.
jetPEI®-Hepatocyte is perfectly suited for plasmid delivery (DNA, shRNA or miRNA) to primary hepatocytes as well as hepatocarcinoma cells, such as Hep G2 and BNL-CL.2.
Every batch of jetPEI®-Hepatocyte is tested in a transfection assay. Typically, transfection of a firefly luciferase gene under the control of the CMV promoter gives 109 RLU (relative light unit)/mg of protein. The value for each batch is indicated on the Certificate of Analysis.
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|Cell Line||in vitro|
|Delivered Molecule||Reagent||Results & Citations|
|Rat primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Ben-Shlomo, S. et al. (2011)
J Hepatol 54, 1214-23
Glucagon-like peptide-1 reduces hepatic lipogenesis via activation of AMP-activated protein kinase
Huang, Y. H. et al. (2014)
Int J Mol Med 34, 677-86
Hydrodynamics-based transfection of rat interleukin-10 gene attenuates porcine serum-induced liver fibrosis in rats by inhibiting the activation of hepatic stellate cells
|Hepatocytes, Huh7||in vitro||miRNA plasmid||jetPEI-Hepatocyte|
Zhang, Y. et al. (2012)
Hepatology 56, 1631-40
Hepatitis C virus-induced up-regulation of microRNA-155 promotes hepatocarcinogenesis by activating Wnt signaling
Andrieux, L. et al. (2004)
Mol Pharmacol 65, 934-43
Aryl hydrocarbon receptor activation and cytochrome P450 1A induction by the mitogen-activated protein kinase inhibitor U0126 in hepatocytes
|Hep G2||in vitro||DNA||jetPEI-Hepatocyte|
Rioux, V. et al. (2013)
J Lipid Res ,
Trans-vaccenate is ∆13-desaturated by FADS3 in rodents
|Human primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Laurenzana, E. M. et al. (2012)
Mol Pharmacol 82, 918-28
The orphan nuclear receptor DAX-1 functions as a potent corepressor of the constitutive androstane receptor (NR1I3)
|COS-1, Rat primary hepatocytes||in vitro||DNA||jetPEI, jetPEI-Hepatocyte|
Baltrusch, S. et al. (2005)
Diabetes 54, 2829-37
Interaction of glucokinase with the liver regulatory protein is conferred by leucine-asparagine motifs of the enzyme
|Rat primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Ezanno, H. et al. (2012)
Lipids 47, 117-28
Myristic acid increases dihydroceramide Delta4-desaturase 1 (DES1) activity in cultured rat hepatocytes
|Hep G2||in vitro||DNA, mRNA||jetPEI-Hepatocyte|
Symens, N. et al. (2012)
Bioconjug Chem 23, 1276-89
Efficient transfection of hepatocytes mediated by mRNA complexed to galactosylated cyclodextrins
|Rat primary hepatocytes||in vitro||shRNA plasmid||jetPEI-Hepatocyte|
Naimi, M. et al. (2007)
Endocrinology 148, 2424-34
Nuclear forkhead box O1 controls and integrates key signaling pathways in hepatocytes