jetPEI®-HUVEC

For endothelial cells

  • Efficient transfection of endothelial cells (up to 50% in HUVEC)
  • As efficient as electroporation
  • Easy-to-use protocol
  • Good cell viability
  • Compatible with serum and antibiotics

Specifications

Reagent

jetPEI®-HUVEC

Molecule delivered

DNA

Applications

DNA transfection of endothelial cells

Cell types

Endothelial cells

Number of transfections

0.5 ml of jetPEI®-HUVEC transfection reagent is sufficient to perform up to 125 transfections in 24-well plates or 40 transfections in 60-mm plates

Storage

4°C, for up to 12 months

Provided with

150 mM NaCl

Summary

jetPEI®-HUVEC is a powerful transfection reagent optimized for the transfection of primary human endothelial cells such as HUVEC (Human umbilical vein endothelial cells). Transfection efficiencies up to 50% have been reached with this reagent.

jetPEI®-HUVEC is also recommended for the transfection of vascular endothelial cells of various origins and appears to be well-suited for such fragile primary cells.

Ordering information

Catalog NumberAmount of reagentAmount of NaCl
108-05N0.5 ml50 ml

0.5 ml of jetPEI®-HUVEC transfection reagent is sufficient to perform up to 125 transfections in 24-well plates

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jetPEI®-HUVEC leads to improved transfection efficiency of endothelial cells

 Compared to jetPEI®, our standard and versatile transfection reagent, jetPEI®-HUVEC provides significantly higher transfection efficiencies in HUVEC cells (Fig. 1).

jetPEI-HUVEC - Comparison jetPEI
Fig. 1: Transfection efficiency and toxicity of jetPEI® and jetPEI®-HUVEC in HUVEC. Cells were incubated for 4 h with transfection complexes in 2 % serum medium. Luciferase activity was measured 24 h after transfection.

Indeed, jetPEI®-HUVEC induces less cellular toxicity than jetPEI®. Using the optimal conditions for jetPEI®-HUVEC, transfection efficiencies of 50% have been reached as observed with a fluorescent protein reporter gene (Fig. 2).

jetPEI-HUVEC - FACS
Fig. 2: FACS analysis of HUVEC cells transfected with a plasmid encoding the EGFP fluorescent protein using jetPEI®-HUVEC. Transfection was performed in 24-well plate with 2 µg of plasmid and 4 µl of jetPEI®-HUVEC (N/P = 5) and in 2 % serum. Complexes were incubated for 4 h with the cells. FACS analysis to quantify EGFP expression was performed 24 h after transfection.

Optimization of the experimental conditions for high transfection efficiency

The effect of serum concentration and incubation time on transfection efficiency was analyzed (Fig. 3).

jetPEI-HUVEC - Incubation with serum
Fig. 3: Effect of low (2 %) versus high (30 %) serum concentrations on transfection efficiency of jetPEI®-HUVEC. HUVEC cells were transfected for 2, 4 or 24 h with 2 µg of pCMVLuc plasmid DNA and 4 µl of jetPEI®-HUVEC in 24-well plates. Luciferase activity was measured 24 h after transfection.

Transfection experiments performed in low (2 %) or high (30 %) concentrations of serum have shown that jetPEI®-HUVEC is most efficient when the cells are grown in low serum concentrations. Gene expression reaches a maximum for HUVEC after 4 h incubation with DNA/jetPEI®-HUVEC complexes.

jetPEI®-HUVEC compares particularly favorably to other non-viral transfection reagents whether cationic lipids or polyamine (Fig. 4).

jetPEI-HUVEC - comparison transfection efficiency
Figure 4: Comparison of transfection efficiency of jetPEI®-HUVEC and other commercially available reagents (P1-P4). Cells were transfected using optimal conditions established for each reagent.

Easy to use protocol

The jetPEI®-HUVEC 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®-HUVEC is compatible with serum and antibiotics, thus eliminating the need for media change. Protein expression is determined 24 h to 72 h post-transfection.

jetPEI®-HUVEC is perfectly suited for plasmid delivery (DNA, shRNA or miRNA plasmids) to fragile cells, such as primary endothelial cells.

Read more…

Here is a selection of relevant references using jetPEI-HUVEC, more are available in our Polyplus-transfection Database.

Feng, J., Zhang, Y., Xing, D. (2012). Low-power laser irradiation (LPLI) promotes VEGF expression and vascular endothelial cell proliferation through the activation of ERK/Sp1 pathway., Cell Signal 24, 1116.

Ganguly, A., Yang, H., Zhang, H., Cabral, F., Patel, K. D. (2013). Microtubule dynamics control tail retraction in migrating vascular endothelial cells., Mol Cancer Ther 12, 2837.

Jin, H. G., Yamashita, H., Nagano, Y., Fukuba, H., Hiji, M., Ohtsuki, T., Takahashi, T., Kohriyama, T., Kaibuchi, K., Matsumoto, M. (2006). Hypoxia-induced upregulation of endothelial small G protein RhoA and Rho-kinase/ROCK2 inhibits eNOS expression., Neurosci Lett408, 62-7.

Wen, H. C., Kao, C., Hsu, R. C., Huo, Y. N., Ting, P. C., Chen, L. C., Hsu, S. P., Juan, S. H., Lee, W. S. (2013). Thy-1-induced migration inhibition in vascular endothelial cells through reducing the RhoA activity., PLoS ONE 8, e6150.

Cristofaro, B., Stone, O. A., Caporali, A., Dawbarn, D., Ieronimakis, N., Reyes, M., Madeddu, P., Bates, D. O., Emanueli, C. (2010). Neurotrophin-3 is a novel angiogenic factor capable of therapeutic neovascularization in a mouse model of limb ischemia., Arterioscler Thromb Vasc Biol30, 1143.

« So far among all the DNA transfection reagents to HUVEC, your jetPEI-HUVEC is the best. The highest efficiency was about 30-50% »

Hong Z, University of Calgary, Canada

Every batch of jetPEI®-HUVEC 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.