Gene transfer and protein dynamics in stem cells using single cell electroporation in a microfluidic device

Share/Save/Bookmark

Valero, A. and Post, J.N. and Nieuwkasteele, J.W. van and Braak, P.M. ter and Kruijer, W. and Berg, A. van den (2008) Gene transfer and protein dynamics in stem cells using single cell electroporation in a microfluidic device. Lab on a Chip, 8 (1). pp. 62-67. ISSN 1473-0197

[img] PDF
Restricted to UT campus only
: Request a copy
2MB
Abstract:There is great interest in genetic modification of bone marrow-derived mesenchymal stem cells (MSC), not only for research purposes but also for use in (autologous) patient-derived-patient-used transplantations. A major drawback of bulk methods for genetic modifications of (stem) cells, like bulk-electroporation, is its limited yield of DNA transfection (typically then 10%). This is even more limited when cells are present at very low numbers, as is the case for stem cells. Here we present an alternative technology to transfect cells with high efficiency (>75%), based on single cell electroporation in a microfluidic device. In a first experiment we show that we can successfully transport propidium iodide (PI) into single mouse myoblastic C2C12 cells. Subsequently, we show the use of this microfluidic device to perform successful electroporation of single mouse myoblastic C2C12 cells and single human MSC with vector DNA encoding a green fluorescent-erk1 fusion protein (EGFP-ERK1 (MAPK3)). Finally, we performed electroporation in combination with live imaging of protein expression and dynamics in response to extracellular stimuli, by fibroblast growth factor (FGF-2). We observed nuclear translocation of EGFP-ERK1 in both cell types within 15 min after FGF-2 stimulation. Due to the successful and promising results, we predict that microfluidic devices can be used for highly efficient small-scale genetic modification of cells, and biological experimentation, offering possibilities to study cellular processes at the single cell level. Future applications might be small-scale production of cells for therapeutic application under controlled conditions.


Item Type:Article
Copyright:© 2008 Royal Society of Chemistry
Faculty:
Science and Technology (TNW)
Electrical Engineering, Mathematics and Computer Science (EEMCS)
Research Group:
Link to this item:http://purl.utwente.nl/publications/62133
Official URL:http://dx.doi.org/10.1039/b713420g
Export this item as:BibTeX
EndNote
HTML Citation
Reference Manager

 

Repository Staff Only: item control page

Metis ID: 250866