Human CD34+ Cell Nucleofector Kit
| Cat. No. | VPA-1003 |
| Optimized Protocol |  Download (PDF, 156 KB) |
| Transfection Efficiency | After 20 hours: 71±11% |
| Cell Database |  CD34+ cell, human |
It is currently under debate if hematopoietic and endothelial cells share common progenitors. CD34+ hematopoietic progenitor cells have recently been shown to differentiate into endothelial precursor cells which are able to induce neovascularization in adults after myocardium ischemia. Simple non-viral transfection of CD34+ cells thus offers new possibilities in cardiovascular research to further investigate the processes of neovascularization with potential future therapeutic applications.
The Nucleofector technology is the first non-viral method for gene transfer into unstimulated CD34+ hematopoietic progenitor cells. As the DNA is directly delivered into the cell's nucleus, very high gene transfer efficiencies can be obtained, equaling those of retroviral systems with up to 70%.
Unstimulated CD34+ cells used for nucleofection can be derived from fresh or cryopreserved cord blood or leukapheresis material. Between 2x105 and 1x107 can be transfected per nucleofection experiment using 1 to 5 μg of plasmid DNA.
Average transfection efficiencies obtained for CD34+ cells.
Partially enriched CD34+ cells were transfected using the Human CD34 Cell Nucleofector Kit and a plasmid encoding the mouse MHC class I heavy chain molecule H-2Kk. Transfection efficiencies were analyzed by flow cytometry 20 hours post nucleofection. Dead cells were excluded from the analysis by propidium iodide staining and gating.
Components of Human CD34 Cell Nucleofector Kit
Cat. No. VPA-1003, 25 Reactions
| 25 | Plastic pipettes | |
| 25 | Certified cuvettes | |
| 10 | µg | pmaxGFP (for 5 reactions) |
| 0.5 | ml | Supplement |
| 2.25 | ml | Human CD34 Nucleofector Solution |
