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Tat peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of progenitor cells

Nature Biotechnology volume 18, pages 410414 (2000) | Download Citation

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Abstract

The ability to track the distribution and differentiation of progenitor and stem cells by high-resolution in vivo imaging techniques would have significant clinical and research implications. We have developed a cell labeling approach using short HIV-Tat peptides to derivatize superparamagnetic nanoparticles. The particles are efficiently internalized into hematopoietic and neural progenitor cells in quantities up to 10–30 pg of superparamagnetic iron per cell. Iron incorporation did not affect cell viability, differentiation, or proliferation of CD34+ cells. Following intravenous injection into immunodeficient mice, 4% of magnetically CD34+ cells homed to bone marrow per gram of tissue, and single cells could be detected by magnetic resonance (MR) imaging in tissue samples. In addition, magnetically labeled cells that had homed to bone marrow could be recovered by magnetic separation columns. Localization and retrieval of cell populations in vivo enable detailed analysis of specific stem cell and organ interactions critical for advancing the therapeutic use of stem cells.

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References

  1. 1.

    The homing of hematopoietic stem cells to the bone marrow. Am. J. Med. Sci. 309, 260–266 (1995).

  2. 2.

    & Homing and trafficking of hemopoietic progenitor cells. Acta Haematol. 97, 97–104 (1997).

  3. 3.

    , , , & Endothelial selectins and vascular cell adhesion molecule-1 promote hematopoietic progenitor homing to bone marrow. Proc. Natl. Acad. Sci. USA 95, 14423–14428 (1998).

  4. 4.

    & Marrow engraftment of hematopoietic stem and progenitor cells is independent of Galphai-coupled chemokine receptors . Exp. Hematol. 27, 946– 955 (1999).

  5. 5.

    et al. Dependence of human stem cell engraftment and repopulation of NOD/SCID mice on CXCR4. Science 283, 845 –848 (1999).

  6. 6.

    , , , & Homing of human cells in the fetal sheep model: modulation by antibodies activating or inhibiting very late activation antigen-4- dependent function. Blood 94, 2515–2522 (1999).

  7. 7.

    et al. Isolation of putative progenitor endothelial cells for angiogenesis . Science 275, 964–967 (1997).

  8. 8.

    , , , & Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors. Science 283, 682–686 (1999).

  9. 9.

    & In situ analysis of lymphocyte migration to lymph nodes. Cell Adhes. Commun. 6, 85–96 (1998).

  10. 10.

    & Scope and perspectives of intravital microscopy-bridge over from in vitro to in vivo. Immunol. Today 14, 519–522 ( 1993).

  11. 11.

    , , , & Homing of fluorescently labeled murine hematopoietic stem cells. Exp. Hematol. 24, 129–140 (1996).

  12. 12.

    , & Hematopoietic stem cell tracking in vivo: a comparison of short-term and long-term repopulating cells. Blood 93, 1916–1921 (1999).

  13. 13.

    , & Constant time imaging approaches to NMR microscopy. Int. J. Imaging Sci. Technol. 8, 263–276 (1997).

  14. 14.

    , , & Looking deeper into vertebrate development. Trends Cell Biol. 9, 73–76 (1999).

  15. 15.

    et al. Histology by magnetic resonance microscopy. Magn. Reson. Q. 9, 1–30 ( 1993).

  16. 16.

    & Use of magnetic techniques for the isolation of cells. J. Chromatogr. B 722, 33–53 (1999).

  17. 17.

    , , & Magnetically labeled cells can be detected by MR imaging. J. Magn. Reson. Imaging 7, 258–263 ( 1997).

  18. 18.

    , , , & Intracellular magnetic labeling of lymphocytes for in vivo trafficking studies. BioTechniques 24, 642–651 (1998).

  19. 19.

    et al. Detection of single mammalian cells by high-resolution magnetic resonance imaging. Biophys. J. 76, 103– 109 (1999).

  20. 20.

    et al. Nuclear magnetic resonance (NMR) imaging of iron oxide-labeled neural transplants. Exp. Neurol. 121, 181 –192 (1993).

  21. 21.

    , , , & MR tracking of magnetically labeled glial cells. Radiol. Soc. North Am. 213, 225 (1999).

  22. 22.

    et al. In vivo MR imaging of transgene expression. Nat. Med. 6, 351–354 ( 2000).

  23. 23.

    , , & In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285, 1569– 1572 (1999).

  24. 24.

    et al. Tat-mediated delivery of heterologous proteins into cells . Proc. Natl. Acad. Sci. USA 91, 664– 668 (1994).

  25. 25.

    et al. Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. Nat. Med. 4, 1449–1452 (1998).

  26. 26.

    , , & High-efficiency intracellular magnetic labeling with novel superparamagnetic Tat peptide conjugates . Bioconjug. Chem. 10, 186– 191 (1999).

  27. 27.

    et al. Cell internalization of the third helix of the Antennapedia homeodomain is receptor-independent. J. Biol. Chem. 271, 18188–18193 (1996).

  28. 28.

    , , , & Polyreactive anti-DNA monoclonal antibodies and a derived peptide as vectors for the intracytoplasmic and intranuclear translocation of macromolecules. Proc. Natl. Acad. Sci. USA 95, 5601–5606 (1998).

  29. 29.

    , & Intercellular delivery of functional p53 by the herpesvirus protein VP22. Nat. Biotechnol. 16, 440– 443 (1998).

  30. 30.

    & Endocytosis and targeting of exogenous HIV-1 Tat protein. EMBO J. 10, 1733– 1739 (1991).

  31. 31.

    et al. Release, uptake, and effects of extracellular human immunodeficiency virus type 1 Tat protein on cell growth and viral transactivation. J. Virol. 67, 277–287 ( 1993).

  32. 32.

    , , , & Cytokine manipulation of primitive human hematopoietic cell self- renewal. Proc. Natl. Acad. Sci. USA 94, 4698–4703 ( 1997).

  33. 33.

    , , & Use of USPIO-induced magnetic susceptibility artifacts to identify sentinel lymph nodes and lymphatic drainage patterns. Magn. Reson. Imaging 16, 917–923 (1998).

  34. 34.

    , , & MR susceptometry: an external-phantom method for measuring bulk susceptibility from field-echo phase reconstruction maps. J. Magn. Reson. Imaging 4, 809–818 (1994).

  35. 35.

    , , , & Monocrystalline iron oxide nanocompounds (MION): physicochemical properties. Magn. Reson. Med. 29, 599–604 (1993).

  36. 36.

    et al. Superparamagnetic iron oxide: pharmacokinetics and toxicity . American Journal of Roentgeneology 152, 167–173 (1989).

  37. 37.

    et al. MR lymphangiography using ultrasmall superparamagnetic iron oxide in patients with primary abdominal and pelvic malignancies. Am. J. Roentgenol. 172, 1347–1351 (1999).

  38. 38.

    , & Neural stem cells as engraftable packaging lines can mediate gene delivery to microglia: evidence from studying retroviral env-related neurodegeneration. J. Virol. 73, 6841– 6851 (1999).

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Acknowledgements

The authors would like to thank Dr. E. Snyder for the kind gift of the neural C17.2 progenitor cells, Drs. N. Michaud and C. Lin for help with the confocal microscopy experiment, Dr. R. Bhorade for performing the uptake experiments in neural progenitor cells, Drs. A. Moore, S. Bredow, and A. Wall for technical assistance, and Dr. L. Garrido for performing some of the original MR imaging studies at 4.7 T. This work was supported in part by RO1 CA59649, RO1 CA46973, RO1 AI/CA 46973 to R.W., RO1 HL55718, RO1 DK50234, DARPA to D.T.S., a development grant from MGH-CMIR, the "Clafin" Distinguished Scholar Award, and Partners (Nesson) Investigator Award to N.C., and the“Bourse Lavoisier” from the French government and the French Society of Radiology to M.L.

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Author notes

    • Maïté Lewin
    •  & Nadia Carlesso

    These authors contributed equally.

Affiliations

  1. Center of Molecular Imaging Research, Massachusetts General Hospital, Charlestown, MA 02129.

    • Maïté Lewin
    • , Ching-Hsuan Tung
    •  & Ralph Weissleder
  2. Experimental Hematology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129 .

    • Nadia Carlesso
    •  & David T. Scadden
  3. Department of Nuclear Engineering, NMR facility, Massachusetts Institute of Technology, Cambridge, MA 02139 .

    • Xiao-Wu Tang
    •  & David Cory

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Correspondence to Ralph Weissleder.

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DOI

https://doi.org/10.1038/74464

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