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OBJECTIVE: Mesenchymal stem-cell (MSC) injection after infarction as regenerative therapy is under examination. The direct visualisation by fluorescent imaging (FI) of these cells and localisation during injection supervise the therapy concept.
METHODS: For in-vivo visualisation of indocyanine-green marced (ICG) MSCs by FI a previously established model of myocardial infarction in pigs was used. MSCs were isoloated from male pigs. Isolation, purification and culture was carried out by standard protocol. MSCs were charcterized using microscopy and flow-cytometry. The MSCs were dyed by incubation with ICG for 12 hours after passage 5. Further cultures of the remaining cells were performed to for control in further passages. In an open-chest model acute transmural myocardial infraction was created in 11 female pigs by occlusion of the left anterior descending artery for one hour. During intracoronary or intramyocardial injection of the dyed cells FI was performed to visualize and detect the distribution of the MSCs.
RESULTS: After 5 passages of MSC, flow cytometry results showed CD45(-), CD14(-), CD90(+), CD105(+), MHCI(+), cells presented similar morphology and oteogenic, chondrocenic and adipogenic differentiation was successful. Incubation with ICG dyed the cells. Further culture and passage of the MSC were possible. Cultured MSCs could be visualized by FI. Intracoronary and intramuscular injection of dyed cells was possible and the visualisation by FI presented excellent pictures for evaluation.
CONCLUSIONS: MSC can be successfully marked with ICG and visualized in-vivo by FI in a porcine myocardial infarct model. Therefore it might be a good diagnostic tool and simple technology for investigation of therapeutic injection of MSCs.