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Self-assembling nanocomplexes by combining ferumoxytol, heparin and protamine for cell tracking by magnetic resonance imaging

Nature Medicine volume 18, pages 463–467 (2012)Cite this article

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Abstract

We report on a new straightforward magnetic cell-labeling approach that combines three US Food and Drug Administration (FDA)-approved drugs—ferumoxytol, heparin and protamine—in serum-free medium to form self-assembling nanocomplexes that effectively label cells for in vivo magnetic resonance imaging (MRI). We observed that the ferumoxytol-heparin-protamine (HPF) nanocomplexes were stable in serum-free cell culture medium. HPF nanocomplexes show a threefold increase in T2 relaxivity compared to ferumoxytol. Electron microscopy showed internalized HPF in endosomes, which we confirmed by Prussian blue staining of labeled cells. There was no long-term effect or toxicity on cellular physiology or function of HPF-labeled hematopoietic stem cells, bone marrow stromal cells, neural stem cells or T cells when compared to controls. In vivo MRI detected 1,000 HPF-labeled cells implanted in rat brains. This HPF labeling method should facilitate the monitoring by MRI of infused or implanted cells in clinical trials.

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Figure 1: Characteristics of self-assembling heparin (H), protamine (P) and ferumoxytol (F) nanocomplexes (HPFs).
Figure 2: Representative light microscopy images of PB-DAB–stained HPF-labeled human stem or immune cells.
Figure 3: Internalization and encapsulation of HPF nanocomplexes in HPF-labeled cells.
Figure 4: Cellular physiological activity in HPF-labeled stem or immune cells.
Figure 5: In vivo magnetic resonance visualization of intracranially implanted HPF-labeled human BMSCs in the rodent brain at 3 T.

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Acknowledgements

This work was supported by and performed in part in the intramural research program at the Clinical Center of the US National Institutes of Health (NIH). BMSCs were provided from the Center for Bone Marrow Stromal Cell Transplantation at the NIH. NSCs were provided as part of a material transfer agreement with K. Aboody (City of Hope Medical Center). The authors would like to acknowledge the Clinical Image Processing Section at the NIH. This work was also supported by NIH grant R01CA122031.

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Authors and Affiliations

  1. Department of Radiology and Imaging Sciences, Frank Laboratory and Laboratory of Diagnostic Radiology Research, US National Institutes of Health (NIH), Bethesda, Maryland, USA

    Mya S Thu, L Henry Bryant, Tiziana Coppola, E Kay Jordan, Matthew D Budde, Bobbi K Lewis, Aneeka Chaudhry & Joseph A Frank

  2. Department of Transfusion Medicine, Cell Processing Section, Clinical Center, NIH, Bethesda, Maryland, USA

    Jiaqiang Ren

  3. Department of Radiology, Cellular and Molecular Imaging Laboratory, Henry Ford Hospital, Detroit, Michigan, USA

    Nadimpalli Ravi S Varma & Ali S Arbab

  4. National Institute of Biomedical Imaging and Bioengineering, NIH, Bethesda, Maryland, USA.,

    Joseph A Frank

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Contributions

J.A.F., M.S.T. and L.H.B. were guarantors of integrity of entire study; J.A.F. and M.S.T. conceptualized and designed the study; M.S.T., L.H.B., E.K.J., B.K.L., A.C., M.D.B., N.R.S.V., J.R., T.C. and A.S.A. performed experiments; J.A.F., M.S.T., L.H.B., E.K.J., J.R. and A.S.A. analyzed and interpreted the data; J.A.F., M.S.T. and L.H.B. drafted the manuscript or revised for important intellectual content; all authors approved the manuscript; M.S.T. performed statistical analysis; J.A.F. and M.S.T. edited the manuscript.

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Correspondence to Joseph A Frank.

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Supplementary Figures 1–5, Supplementary Table 1 and Supplementary Methods (PDF 14869 kb)

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Thu, M., Bryant, L., Coppola, T. et al. Self-assembling nanocomplexes by combining ferumoxytol, heparin and protamine for cell tracking by magnetic resonance imaging. Nat Med 18, 463–467 (2012). https://doi.org/10.1038/nm.2666

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