Aptamer-based isolation and subsequent imaging of mesenchymal stem cells in ischemic myocard by magnetic resonance imaging.
OBJECTIVE
Mesenchymal stem cells (MSC) seem to be a promising cell source for cellular cardiomyoplasty. We recently developed a new aptamer-based specific selection of MSC to provide "ready to transplant" cells directly after isolation. We evaluated MRI tracking of newly isolated and freshly transplanted MSC in the heart using one short ex vivo selection step combining specific aptamer-based isolation and labeling of the cells.


METHODS
Bone marrow (BM) was collected from healthy pigs. The animals were euthanized and the heart was placed in a perfusion model. During cold ischemia, immunomagnetic isolation of MSC from the BM by MSC-specific aptamers labeled with Dynabeads was performed within 2 h. For histological identification the cells were additionally stained with PKH26. Approx. 3 x 10(6) of the freshly aptamer-isolated cells were injected into the ramus interventricularis anterior (RIVA) and 5 x 10(5) cells were injected directly into myocardial tissue after damaging the respective area by freezing (cryo-scar). 3 x 10(6) of the aptamer-isolated cells were kept for further characterization (FACS and differentiation assays). 20 h after cell transplantation, MRI of the heart using a clinical 3.0 Tesla whole body scanner (Magnetom Trio, Siemens, Germany) was performed followed by histological examinations.


RESULTS
The average yield of sorted cells from 120 ml BM was 7 x 10(6) cells. The cells were cultured and showed MSC-like properties. MRI showed reproducible artifacts within the RIVA-perfusion area and the cryo-scar with surprisingly excellent quality. The histological examination of the biopsies showed PKH26-positive cells within the areas which were positive in the MRI in contrast to the control biopsies.


CONCLUSIONS
Immunomagnetic separation of MSC by specific aptamers linked to magnetic particles is feasible, effective and combines a specific separation and labeling technique to a "one stop shop" strategy.
Schäfer
R
R
Institut für Klinische und Experimentelle Transfusionsmedizin, Universitätsklinikum Tübingen.
Wiskirchen
J
J
Guo
K
K
Neumann
B
B
Kehlbach
R
R
Pintaske
J
J
Voth
V
V
Walker
T
T
Scheule
A M
AM
Greiner
T O
TO
Hermanutz-Klein
U
U
Claussen
C D
CD
Northoff
H
H
Ziemer
G
G
Wendel
H P
HP
D000818:Animals; D052157:Aptamers, Nucleotide; D001854:Bone Marrow Cells; D018421:Cardiomyoplasty / Q000379:methods; D002469:Cell Separation; D005240:Feasibility Studies; D005456:Fluorescent Dyes; D018189:Immunomagnetic Separation; D008279:Magnetic Resonance Imaging; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology; D017202:Myocardial Ischemia; D009930:Organic Chemicals; D013194:Staining and Labeling; D013552:Swine; D013997:Time Factors
D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26