Human Dendritic Cell Nucleofector Kit I
(for short-term expression)*
| Cat. No. | VPA-1004 |
| Optimized Protocol |  Download (PDF, 157 KB) |
| Transfection Efficiency | After 24 hours: 49±13% |
| Cell Database |  Dendritic cell, human |
A rapidly growing interest in dendritic cells as both research and therapeutic tools reflects an increased understanding and appreciation of their role at the heart of the immune system and the broad range of functions they perform. Human dendritic cells can be transfected using the same nucleofection conditions with different substrates such as RNA, DNA or siRNA. Nucleofected
dendritic cells usually maintain transgene expression for about 48 hours.
- Transfection efficiencies up to 50%
- For immature and mature monocyte-derived dendritic cells
- For short term expression of up to 48 hours
Nucleofection of human dendritic cells.
Human dendritic cells were differentiated from monocytes using 100ng/ml IL-4 and 50 ng/ml GM-CSF. On day 6 cells were transfected with 5 μg plasmid DNA encoding the green fluorescent protein (eGFP) using the Human Dendritic Cell Nucleofector Kit I. 24 hours post nucleofection cells were analyzed by flow cytometry (FACS). FACS data show mock transfected cells (plasmid without insert) (A) and eGFP transfected cells (B).
(Courtesy of Dr. Esen and Prof. Radbruch
Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), Germany).
Nucleofection of primary dendritic cells shows the role of small GTPase Rho in the regulation of cytoarchitecture.
The GTPase Rho is involved in the control of actin and actin associated proteins. Mature monocyte-derived DC were mock transfected (Mock), or transfected with an empty GFP-encoding vector (vector-GFP) or with a dominant negative form of Rho (N19-Rho). 18h post nucleofection cells were subjected to immunofluorescence. Cells were stained with Cy3-phalloidin to detect F-actin. N19-Rho transfected cells showed actin appearing as a thin ring surrounding the cell periphery and a retraction in the length of the dendrites compared to the control transfections. This result suggests the requirement of Rho in the regulation of dendrite formation and maintenance of mature DC morphology.
(Data taken from Rodriguez-Fernandez et al. (2003) amaxa application note).
Components of Human Dendritic Cell Nucleofector Kit I (for short-term expression)
Cat. No. VPA-1004, 25 Reactions
| 25 | Plastic pipettes | |
| 25 | Certified cuvettes | |
| 10 | µg | pmaxGFP (for 5 reactions) |
| 0.5 | ml | Supplement |
| 2.25 | ml | Human Dendritic Cell Nucleofector Solution |
