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
Store jetPEI®-Hepatocyte at -20°C ± 5°C.
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|
|101000038||jetPEI®-Hepatocyte 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|
Andrieux, L. O. et al. (2007)
Cancer Res 67, 2114-23
GATA-1 is essential in EGF-mediated induction of nucleotide excision repair activity and ERCC1 expression through ERK2 in human hepatoma cells
Arima, H. et al. (2012)
Pharmaceutics 4, 130-48
Polyamidoamine Dendrimer Conjugates with Cyclodextrins as Novel Carriers for DNA, shRNA and siRNA
|Hep G2||in vitro||DNA||jetPEI-Hepatocyte|
Makkonen, K. E. et al. (2013)
J Virol 87, 11148-59
6-o- and N-sulfated syndecan-1 promotes baculovirus binding and entry into Mammalian cells
|Hep G2||in vitro||DNA||jetPEI-Hepatocyte|
Benoki, S. et al. (2012)
Arch Biochem Biophys 517, 123-30
Transactivation of ABCG2 through a novel cis-element in the distal promoter by constitutive androstane receptor but not pregnane X receptor in human hepatocytes
|Hep3B, PLC/PRF/5||in vitro||DNA||jetPEI-Hepatocyte|
Fu, X. et al. (2018)
Mol Cancer 17, 73
Linc00210 drives Wnt/beta-catenin signaling activation and liver tumor progression through CTNNBIP1-dependent manner
|Human primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Zhang, J. et al. (2018)
Cell Host Microbe 23, 819-831 e5
Flaviviruses Exploit the Lipid Droplet Protein AUP1 to Trigger Lipophagy and Drive Virus Production
|COS-7, Hep3B||in vitro||DNA||jetPEI, jetPEI-Hepatocyte|
Takahashi, E. et al. (2014)
Am J Physiol Cell Physiol 306, C334-42
Anaerobic respiration sustains mitochondrial membrane potential in a prolyl hydroxylase pathway-activated cancer cell line in a hypoxic microenvironment
|Human primary hepatocytes, Mouse primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Kim, D. H. et al. (2018)
Nat Commun 9, 3284
Intracellular interleukin-32gamma mediates antiviral activity of cytokines against hepatitis B virus
|Hep G2, Huh7, PLC/PRF/5||in vitro||DNA||jetPEI-Hepatocyte|
Krohler T. et al. (2019)
Cancers 11, 1754
The mRNA-binding Protein TTP/ZFP36 in Hepatocarcinogenesis and Hepatocellular Carcinoma
|Rat primary hepatocytes||in vitro||DNA||jetPEI-Hepatocyte|
Arden, C. et al. (2007)
Diabetes 56, 1773-82
Cell biology assessment of glucokinase mutations V62M and G72R in pancreatic beta-cells: evidence for cellular instability of catalytic activity