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KMID : 1377020150120050371
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Tissue Engineering and Regenerative Medicine
2015 Volume.12 No. 5 p.371 ~ p.378
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Direct labeling of 19F-perfluorocarbon onto multilayered cell sheet for MRI-based non-invasive cell tracking
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Joan Oliva
Fawzia Bardag-Gorce
Andrew Wood
Hiroyuki Sota
Yutaka Niihara
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Abstract
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Autologous stem cell transplantation for eye diseases is immunologically preferable to avoid allograft rejection. However, the fate of the grafted cells has never been studied. Here, we propose to use 19F-perfluorocarbon magnetic resonance imaging tracer agent, to label cell sheet in vitro. This labeling enables non-invasive visualization of possible migration of grafted cells. Oral mucosal epithelial cells were isolated from rabbit oral mucosal epithelium and were cultivated in a thermo-responsive surface to engineer a multilayer cell sheet. Different concentrations of 19F-perfluorocarbon were added to the cell sheet culture media, one or two times. Cells were analyzed in a 7 T nuclear magnetic resonance to determine the labeling efficiency. We found that 10 mg/mL and two incubations with 19F-perfluorocarbon were the optimal condition for labeling. H&E and immunocytochemistry showed that labeling did not affect the expression of cell sheets specific markers (CK4, CK13, connexin43, E-cadherin). Furthermore, no significant effects were observed on the number of cells and the cell viability, making 19F-perfluorocarbon suitable for cell tracking, with no side effects.
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KEYWORD
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Oral mucosa epithelial, Cell sheet, Nuclear magnetic resonance, 19F-perfluorocarbon
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