For the evaluation of new stem cell therapies it is important to non-invasively track the injected cells in vivo. This video will show you how to label human mesenchymal and embryonic stem cells with iron oxide based contrast agents in vivo for subsequent MR imaging in vivo.
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.
Material Name | Type | Company | Catalogue Number | Comment |
---|---|---|---|---|
D-MEM High Glucose | Reagent | Sigma | D5648 | |
Gelatin | Reagent | Sigma | G1890 | dilute Gelatin in PBS to final concentration of 0.1 % |
PBS (Ca++ Mg++ freed) | Reagent | GIBCO | 14190-144 | |
Trypsin-EDTA 0.05% | Reagent | Invitrogen | 25300-120 | |
Trypsin-EDTA 0.25% | Reagent | Invitrogen | 25200-114 | |
FBS characterized | Reagent | Hyclone | SH30071.03 | |
Cell Strainer (40 μm Nylon) | Tool | BD Falcon | 352340 | |
Knockout DMEM | Reagent | GIBCO | 10829018 | |
Knockout Serum Replacer | Reagent | GIBCO | 10828028 | |
b-Mercaptoethanol | Reagent | Sigma | 7522 | |
non-essential amino acids | Reagent | GIBCO | 11140050 | |
FGF-2 | Reagent | R&D Systems | 233-FB-025 | |
Glutamine | Reagent | GIBCO | 25030081 | |
Penicillin/Streptomycin | Reagent | GIBCO | 15140-122 | |
Ferumoxides (Endorem) | Reagent | Guerbet | ||
Ferucarbotran (Resovist®) | Reagent | Bayer Schering AG |