Homing of 111In-Labeled Bone Marrow Mesenchymal Stem Cells in Acute Brain Trauma Model
급성 brain trauma 모델에서 111In으로 표지된 중간엽 줄기세포의 귀소성
This study was to evaluate the in vivo distribution of intravenously transplanted bone marrow-derived mesenchymal stem cells (BMSCs) in an acute brain trauma model by 111In-tropolone labeling and to perform the effect of 111In-labeling on the viability and functions of BMSCs. Rat BMSCs were labeled with 37 MBq 111In-tropolone. Their labeling efficiency and in vitro retention rate were measured. To evaluate dose-dependent effect of 111In-labeling, BMSCs were labeled with various doses (0.4-11.1 Bq/cell) of 111In-tropolone, and growth curve analysis, fluorescent activated cell sorter (FACS) analysis after staining with 5-bromo-2-deoxy-uridine (BrdU), and microscopic evaluation after 5-bromo-4-chloro-3-indolyl-D-galactopyranoside (X-gal) staining were performed until the 14th day. FACS analysis after staining with Annexin V- fluorescein isothiocyanate (FITC) and propidium iodide (PI) was performed at early (3 and 12 hr) and late (7 days) stages with higher doses of 111In (11.1 and 33.3 Bq/cell) to evaluate apoptotic or necrotic change of labeled BMSCs. The biodistribution of 111In-BMSCs in trauma models was compared with those in sham-operated rats and normal rats by gamma camera images. The migration of 111In-BMSCs to the traumatic brain was evaluated using confocal microscope. The labeling efficiency of 111In-BMSCs was 66 ± 5%, and their retention rate was 85.3% at 1 h after labeling. There was no difference in the number of viable cells between 111In-BMSCs and controls at 48 h after labeling. However, the proliferation of 111In-BMSCs was inhibited after the third day of labeling, and it did not reach confluency. For lower doses of 111In (0.4 and 1.1 Bq/cell), the growth of labeled stem cells was not significantly different from that of control, whereas, labeling with higher doses of 111In (4.4 and 11.1 Bq/cell) led to a significant proliferative inhibition from the 3rd day to the 14th day. FACS analysis also revealed less BrdU positive cells in BMSCs labeled with 1.1, 4.4 and 11.1 Bq/cell compared with controls on the 3rd day after labeling. Of these, the patterns of cell cycle in BMSCs labeled with 0.4 and 1.1 Bq/cell of 111In were restored similar to controls on the 14th day. On the contrary, BMSCs labeled with 4.4 and 11.1 Bq/cell of 111In could not recover from cell cycle arrest. Senescence-associated β-gal (SA- β-gal) staining was not prominent in all concentrations until the 14th day after labeling. FACS analysis with Annexin V-FITC and PI also revealed no significant apoptosis or necrosis in both early and late stages. On gamma camera images, most of the 111In-BMSCs uptake was observed in the liver and spleen at the second day of injection. The brain uptake of 111In-BMSCs was more prominent in trauma models (1.4%) than in sham-operated (0.5%) or normal rats (0.3%). Radiolabeled BMSCs were observed at the marginal region of traumatic brain on the confocal microscope. We observed the dose-dependent growth inhibition of BMSCs by 111In-labeling, which was developed by dose-dependent, transient cell cycle arrest, not by cellular senescence or apoptosis/necrosis. Although growth inhibition by 111In-labeling need to be evaluated further prior to use in humans, 111In-BMSCs are useful for the tracking of intravenously transplanted mesenchymal stem cells in brain disease models.
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