Contrast Agent Research Group at the Center for Molecular and Functional Imaging, Department of Radiology, University of California San Francisco
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Henning, T. D., Boddington, S., Daldrup-Link, H. E. Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging. J. Vis. Exp. (13), e685, doi:10.3791/685 (2008).
Labeling of hMSC with Ferucarbotran
This is the technique for labeling human mesenchymal stem cells with Ferucarbotran, an iron oxide based contrast agent for MR imaging:
Labeling of human embryonic stem cells with Ferumoxides
A non-invasive depiction of stem cells is crucial for monitoring virtually any stem cell based therapy. A better understanding of the in vivo kinetics of labeled stem cells, as visualized on MR images, could lead the way to a rational and a more effective use of stem cell-based therapies. Since we use clinically applicable contrast agents and imaging techniques, our presented protocols should be readily translatable to applications in patients. Potential clinical applications of our methods include comparative investigations of the engraftment process of various stem cell subtypes or genetically engineered stem cells, as well as, the assessment of therapy effects on the outcome of stem cell engraftment. Results should be immediately helpful in preclinical assessments of stem-based therapies, in the design of related clinical trials, and later, in the assessment of those stem cell-based therapies in clinical practice. Of note, our imaging technique is also applicable to other stem cell populations, such as hematopoietic stem cells or neuronal stem cells.
This project was supported by a Leon J. Thal SEED grant from the California Institute for Regenerative Medicine. Tobias Henning was funded by a Research stipend from the German Research Association (DFG, HE 4578/1-2). We want to gratefully acknowledge Juanito Meneses for his advice on the culture of human embryonic stem cells.
|D-MEM High Glucose||Reagent||Sigma-Aldrich||D5648|
|Gelatin||Reagent||Sigma-Aldrich||G1890||dilute Gelatin in PBS to final concentration of 0.1 %|
|PBS (Ca++ Mg++ freed)||Reagent||GIBCO, by Life Technologies||14190-144|
|Cell Strainer (40 µm Nylon)||Tool||BD Biosciences||352340|
|Knockout DMEM||Reagent||GIBCO, by Life Technologies||10829018|
|Knockout Serum Replacer||Reagent||GIBCO, by Life Technologies||10828028|
|non-essential amino acids||Reagent||GIBCO, by Life Technologies||11140050|
|Glutamine||Reagent||GIBCO, by Life Technologies||25030081|
|Penicillin/Streptomycin||Reagent||GIBCO, by Life Technologies||15140-122|
|Ferucarbotran (ResovistĂ‚®)||Reagent||Bayer AG|
1. Daldrup-Link, H.E., Rudelius, M., Oostendorp, R.A.J., Settles, M., Piontek, G., Metz, S., Heinzmann, U., Rummeny, E.J., Schlegel, J., Link, T.M. Targeting of Hematopoietic Progenitor Cells with MR Contrast Agents. Radiology 228, 760-767 (2003)
2. Daldrup-Link, H.E., Rudelius, M., Piontek, G., Metz, S., Bräuer, R., Debus, G., Corot, C., Schlegel, J., Link, T.M., Peschel, C., Rummeny, E.J., Oostendorp, R.A.J. Migration of iron oxide labeled human hematopoietic progenitor cells in a xenotransplant model: in vivo monitoring using clinical magnetic resonance imaging equipment. Radiology 234, 197-205 (2005)
3. Henning, T.D., Saborowski, O., Golovko, D., Boddington, S., Bauer, J.S., Fu, Y., Meier, R., Pietsch, H., Sennino, B., McDonald, D.M., Daldrup-Link, H.E. Cell labeling with the positive MR contrast agent Gadofluorine M. Eur Radiol 17, 1226-34 (2007)