Overview
Specifications
| Reagent | jetPEI®-Hepatocyte |
|---|---|
| Molecule delivered | DNA |
| Applications | DNA transfection of primary hepatocytes |
| Cell types | 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 |
| Storage | 5°C ± 3°C, for up to 12 months |
| Provided with | 150 mM NaCl |
Summary
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.
Ordering information
| Reference Number | Amount of reagent | Amount of NaCl |
|---|---|---|
| 102-05N | 0.5 ml | 50 ml |
Description
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.
Applications
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.
Quality
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.
Bibliography
| Cell Line | in vitro in vivo | Delivered Molecule | Reagent | Results & Citations | |
|---|---|---|---|---|---|
| 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) | More details |
| B16A2 | in vitro | DNA | jetPEI-Hepatocyte | 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 | More details |
| Hep G2 | in vitro | DNA | jetPEI-Hepatocyte | Rioux, V. et al. (2013) J Lipid Res , Trans-vaccenate is ∆13-desaturated by FADS3 in rodents | More details |
| 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 | More details |
| 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 | More details |
| 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 | More details |
| Hep G2 | in vitro | DNA | jetPEI-Hepatocyte | Polishchuk, E. V. et al. (2014) Dev Cell 29, 686-700 Wilson disease protein ATP7B utilizes lysosomal exocytosis to maintain copper homeostasis | More details |
| 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 | More details |
| Hep G2, PLC/PRF/5 | in vitro | DNA, siRNA | INTERFERin, jetPEI-Hepatocyte | Kessler, S. M. et al. (2013) Am J Physiol Gastrointest Liver Physiol 304, G328-36 IGF2 mRNA binding protein p62/IMP2-2 in hepatocellular carcinoma: antiapoptotic action is independent of IGF2/PI3K signaling | More details |
| 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 | More details |