Molecular imaging of endothelial progenitor cell engraftment using contrast-enhanced ultrasound and targeted microbubbles

Michael A. Kuliszewski, Hiroko Fujii, Christine Liao, Alexandra H. Smith, Aris Xie, Jonathan Lindner, Howard Leong-Poi

    Research output: Contribution to journalArticle

    40 Citations (Scopus)

    Abstract

    Aims: Imaging methods to track the fate of progenitor cells after their delivery would be useful in assessing the efficacy of cell-based therapies. We hypothesized that contrast-enhanced ultrasound (CEU) using microbubbles targeted to a genetically engineered cell-surface marker on endothelial progenitor cells (EPCs) would allow the targeted imaging of vascular engraftment. Methods and results: Rodent bone marrow-derived EPCs were isolated, cultured, and transfected to express the marker protein, H-2Kk, on the cell surface. Non-transfected EPCs and EPCs transfected with either null plasmid or Firefly luciferase served as controls. Control microbubbles (MBC) and microbubbles targeted to H-2Kk expressed on EPCs (MBH-2Kk) were constructed. Binding of targeted microbubbles to EPCs was assessed in vitro using a parallel plate flow chamber system. CEU imaging of EPC-targeted microbubbles was assessed in vivo using subcutaneously implanted EPC-supplemented Matrigel plugs in rats. In flow chamber experiments, there was minimal attachment of microbubbles to plated control EPCs. Although numbers of adhered MBC were also low, there was greater and more diffuse attachment of MBH-2Kk to plated H-2Kk-transfected EPCs. Targeted CEU demonstrated marked contrast enhancement at the periphery of the H-2Kk-transfected EPC-supplemented Matrigel plug for MBH-2Kk, whereas contrast enhancement was low for MBC. Contrast enhancement was also low for both microbubbles within control mock-transfected EPC plugs. The signal intensity within the H-2Kk-transfected EPC plug was significantly greater for MBH-2Kk when compared with MBC. Conclusion: Microbubbles targeted to a genetically engineered cell-surface marker on EPCs exhibit specific binding to EPCs in vitro. These targeted microbubbles bind to engrafted EPCs in vivo within Matrigel plugs and can be detected by their enhancement on CEU imaging.

    Original languageEnglish (US)
    Pages (from-to)653-662
    Number of pages10
    JournalCardiovascular Research
    Volume83
    Issue number4
    DOIs
    StatePublished - Sep 2009

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    Microbubbles
    Molecular Imaging
    Endothelial Progenitor Cells
    Ultrasonography
    Firefly Luciferases
    Cell- and Tissue-Based Therapy

    Keywords

    • Contrast ultrasound
    • Endothelial progenitor cells
    • Molecular imaging

    ASJC Scopus subject areas

    • Cardiology and Cardiovascular Medicine
    • Physiology (medical)
    • Physiology

    Cite this

    Molecular imaging of endothelial progenitor cell engraftment using contrast-enhanced ultrasound and targeted microbubbles. / Kuliszewski, Michael A.; Fujii, Hiroko; Liao, Christine; Smith, Alexandra H.; Xie, Aris; Lindner, Jonathan; Leong-Poi, Howard.

    In: Cardiovascular Research, Vol. 83, No. 4, 09.2009, p. 653-662.

    Research output: Contribution to journalArticle

    Kuliszewski, Michael A. ; Fujii, Hiroko ; Liao, Christine ; Smith, Alexandra H. ; Xie, Aris ; Lindner, Jonathan ; Leong-Poi, Howard. / Molecular imaging of endothelial progenitor cell engraftment using contrast-enhanced ultrasound and targeted microbubbles. In: Cardiovascular Research. 2009 ; Vol. 83, No. 4. pp. 653-662.
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    AU - Fujii, Hiroko

    AU - Liao, Christine

    AU - Smith, Alexandra H.

    AU - Xie, Aris

    AU - Lindner, Jonathan

    AU - Leong-Poi, Howard

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    N2 - Aims: Imaging methods to track the fate of progenitor cells after their delivery would be useful in assessing the efficacy of cell-based therapies. We hypothesized that contrast-enhanced ultrasound (CEU) using microbubbles targeted to a genetically engineered cell-surface marker on endothelial progenitor cells (EPCs) would allow the targeted imaging of vascular engraftment. Methods and results: Rodent bone marrow-derived EPCs were isolated, cultured, and transfected to express the marker protein, H-2Kk, on the cell surface. Non-transfected EPCs and EPCs transfected with either null plasmid or Firefly luciferase served as controls. Control microbubbles (MBC) and microbubbles targeted to H-2Kk expressed on EPCs (MBH-2Kk) were constructed. Binding of targeted microbubbles to EPCs was assessed in vitro using a parallel plate flow chamber system. CEU imaging of EPC-targeted microbubbles was assessed in vivo using subcutaneously implanted EPC-supplemented Matrigel plugs in rats. In flow chamber experiments, there was minimal attachment of microbubbles to plated control EPCs. Although numbers of adhered MBC were also low, there was greater and more diffuse attachment of MBH-2Kk to plated H-2Kk-transfected EPCs. Targeted CEU demonstrated marked contrast enhancement at the periphery of the H-2Kk-transfected EPC-supplemented Matrigel plug for MBH-2Kk, whereas contrast enhancement was low for MBC. Contrast enhancement was also low for both microbubbles within control mock-transfected EPC plugs. The signal intensity within the H-2Kk-transfected EPC plug was significantly greater for MBH-2Kk when compared with MBC. Conclusion: Microbubbles targeted to a genetically engineered cell-surface marker on EPCs exhibit specific binding to EPCs in vitro. These targeted microbubbles bind to engrafted EPCs in vivo within Matrigel plugs and can be detected by their enhancement on CEU imaging.

    AB - Aims: Imaging methods to track the fate of progenitor cells after their delivery would be useful in assessing the efficacy of cell-based therapies. We hypothesized that contrast-enhanced ultrasound (CEU) using microbubbles targeted to a genetically engineered cell-surface marker on endothelial progenitor cells (EPCs) would allow the targeted imaging of vascular engraftment. Methods and results: Rodent bone marrow-derived EPCs were isolated, cultured, and transfected to express the marker protein, H-2Kk, on the cell surface. Non-transfected EPCs and EPCs transfected with either null plasmid or Firefly luciferase served as controls. Control microbubbles (MBC) and microbubbles targeted to H-2Kk expressed on EPCs (MBH-2Kk) were constructed. Binding of targeted microbubbles to EPCs was assessed in vitro using a parallel plate flow chamber system. CEU imaging of EPC-targeted microbubbles was assessed in vivo using subcutaneously implanted EPC-supplemented Matrigel plugs in rats. In flow chamber experiments, there was minimal attachment of microbubbles to plated control EPCs. Although numbers of adhered MBC were also low, there was greater and more diffuse attachment of MBH-2Kk to plated H-2Kk-transfected EPCs. Targeted CEU demonstrated marked contrast enhancement at the periphery of the H-2Kk-transfected EPC-supplemented Matrigel plug for MBH-2Kk, whereas contrast enhancement was low for MBC. Contrast enhancement was also low for both microbubbles within control mock-transfected EPC plugs. The signal intensity within the H-2Kk-transfected EPC plug was significantly greater for MBH-2Kk when compared with MBC. Conclusion: Microbubbles targeted to a genetically engineered cell-surface marker on EPCs exhibit specific binding to EPCs in vitro. These targeted microbubbles bind to engrafted EPCs in vivo within Matrigel plugs and can be detected by their enhancement on CEU imaging.

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