Viability and proliferation potential of adipose-derived stem cells following labeling with a positron-emitting radiotracer
- 307 Downloads
Adipose-derived stem cells (ASCs) have promising potential in regenerative medicine and cell therapy. Our objective is to examine the biological function of the labeled stem cells following labeling with a readily available positron emission tomography (PET) tracer, 18F-fluoro-2-deoxy-D-glucose (FDG). In this work we characterize labeling efficiency through assessment of FDG uptake and retention by the ASCs and the effect of FDG on cell viability, proliferation, transdifferentiation, and cell function in vitro using rat ASCs.
Samples of 105 ASCs (from visceral fat tissue) were labeled with concentrations of FDG (1–55 Bq/cell) in 0.75 ml culture medium. Label uptake and retention, as a function of labeling time, FDG concentration, and efflux period were measured to determine optimum cell labeling conditions. Cell viability, proliferation, DNA structure damage, cell differentiation, and other cell functions were examined. Non-labeled ASC samples were used as a control for all experimental groups. Labeled ASCs were injected via tail vein in several healthy rats and initial cell biodistribution was assessed.
Our results showed that FDG uptake and retention by the stem cells did not depend on FDG concentration but on labeling and efflux periods and glucose content of the labeling and efflux media. Cell viability, transdifferentiation, and cell function were not greatly affected. DNA damage due to FDG radioactivity was acute, but reversible; cells managed to repair the damage and continue with cell cycles. Over all, FDG (up to 25 Bq/cell) did not impose severe cytotoxicity in rat ASCs. Initial biodistribution of the FDG-labeled ASCs was 80% + retention in the lungs. In the delayed whole-body images (2–3 h postinjection) there was some activity distribution resembling typical FDG uptake patterns.
For in vivo cell tracking studies with PET tracers, the parameter of interest is the amount of radiotracer that is present in the cells being labeled and consequent biological effects. From our study we developed a labeling protocol for labeling ASCs with a readily available PET tracer, FDG. Our results indicate that ASCs can be safely labeled with FDG concentration up to 25 Bq/cell, without compromising their biological function. A labeling period of 90 min in glucose-free medium and efflux of 60 min in complete media resulted in optimum label retention, i.e., 60% + by the stem cells. The initial biodistribution of the implanted FDG-labeled stem cells can be monitored using microPET imaging.
KeywordsCell labeling Adipose-derived stem cells 18F-FDG PET Cell tracking
Adipose-derived stem cells
- 18F-FDG, FDG
Positron emission tomography
The authors thank the reviewers of this manuscript for their insightful comments and suggestions. This work has been supported by a Canadian Institute of Health Research (CIHR) Emerging Team Grant in Regenerative Medicine and Nanomedicine.
Conflicts of interest
- 6.Thomson M, Wall DM, Hicks RJ, Prince HM. In vivo tracking for cell therapies. Q J Nucl Med Mol Imaging 2005;49:339–48.Google Scholar
- 8.Terrovitis J, Lautamäki R, Bonios M, Fox J, Engles JM, Yu J, et al. Noninvasive quantification and optimization of acute cell retention by in vivo positron emission tomography after intramyocardial cardiac-derived stem cell delivery. J Am Coll Cardiol 2009;54(17):1619–26.PubMedCrossRefGoogle Scholar
- 12.Vesselle H, Grierson J, Muzi M, Pugsley JM, Schmidt RA, Rabinowitz P, et al. In vivo validation of 3′-deoxy-3′-[18F]fluorothymidine ([18F]FLT) as a proliferation imaging tracer in humans: correlation of [(18)F]FLT uptake by positron emission tomography with Ki-67 immunohistochemistry and flow cytometry in human lung tumors. Clin Cancer Res 2002;8(11):3315–23.PubMedGoogle Scholar
- 16.Wang L, Deng J, Wang J, Xiang B, Yang T, Gruwel M, et al. Superparamagnetic iron oxide does not affect the viability and function of adipose-derived stem cells, and superparamagnetic iron oxide-enhanced magnetic resonance imaging identifies viable cells. Magn Reson Imaging 2009;27:108–19.PubMedCrossRefGoogle Scholar
- 21.Hall EJ. Radiobiology for the radiologist. 4th ed. Philadelphia: Lippincott; 1994.Google Scholar