Efficient magnetic cell labeling with protamine sulfate complexed to ferumoxides for cellular MRI

Arbab, Ali S.; Yocum, Gene T.; Kalish, Heather; Jordan, Elaine; Anderson, Stasia A.; Khakoo, Aarif Y.; Read, Elizabeth J.; Frank, Joseph A.

doi:10.1182/blood-2004-02-0655

Cited by 515 publications

(512 citation statements)

References 26 publications

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“…Results revealed a positive induction within 7 days, demonstrated by staining of cartilage-specific proteogylcans. A mean cellular iron content of 24 pg was found in co-labeled MSC, this finding was similar to reported values for magnetic cell labeling without toxic effects [10,41,43]. Therefore, it was assumed that the co-labeling approach described above was an applicable method to study MSC in-vivo by MRI and to validate their localization by histology.…”

Section: Discussionsupporting

confidence: 82%

“…Another important issue was the differentiation capacity of co-labeled MSC. Different studies have shown that osteogenic and adipogenic differentiation of MSC is not affected by magnetic labeling, but considering contradictory results for chondrogenic differentiation [41,42], this seems to be sensitive to cellular iron uptake. Therefore, the chondrogenic differentiation capacity was investigated for co-labeled MSC.…”

Section: Discussionmentioning

confidence: 95%

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Magnetic Resonance Imaging of Single Co-Labeled Mesenchymal Stromal Cells after Intracardial Injection in Mice

Salamon

Wicklein

Didié

et al. 2014

Fortschr Röntgenstr

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Ziel: Etablierung einer Doppelmarkierung mesenchymaler Stromazellen (MSZ) zur in-vivo Detektion einzelner MSZ mittels MRT und zur histologischen Validierung. Material und Methoden: Murine MSZ wurden mit fluoreszierenden Eisenmikropartikeln und Carboxyfluorescein Succinimidyl Ester (CFSE) markiert. Der zelluläre Eisengehalt wurde mittels Atomabsorptionsspektrometrie bestimmt. Zellprolieferation und Expression charakteristischer Oberflächenmarker wurden mittels Durchflusszytometrie bestimmt. Die chondrogene Differenzierungskapazität wurde überprüft. Verschiedene Zellanzahlen (n1 = 5000, n2 = 15 000, n3 = 50 000) wurden bei 12 Mäusen in den linken Herzventrikel injiziert. Es erfolgte die sequenzielle MRT der Tiere an einem klinischen 3.0 T MRT. Zur histologischen Validierung wurden Kryostatschnitte fluoreszensmikroskopisch untersucht. Ergebnisse: Die magnetische und fluoreszierende Doppelmarkierung von MSZ wurde etabliert (mittlerer zellulärer Eisengehalt 23,6 ± 4,3 pg). Durchflusszytometrisch zeigten sich ähn-liche Zellprolieferationsraten und Rezeptorexpressionsprofile von markierten und unmarkierten MSZ. Die chondrogene Differenzierung der doppelt markierten MSZ wurde verifiziert. Nach Zellinjektion zeigten sich im MRT multiple Signalauslöschungen im Hirn und geringer in der Niere. Im Hirn fanden sich durchschnittlich 4,6 ± 1,2 (n1), 9,0 ± 3,6 (n2) und 25,0 ± 1,0 (n3) Signalauslöschungen pro Schicht. Durchschnittlich fanden sich 8,7 ± 3,1 (n1), 22,0 ± 6,1 (n2) und 89,8 ± 6,5 (n3) Zellen pro korrespondierendem Kryostatschnitt. Die statistische Korrelation der mittels MRT detektierten Signalauslöschungen und der histologisch nachgewiesenen Zellen ergab einen Korrelationskoeffizienten von r = 0,91. Schlussfolgerung: Die Studie zeigt die erfolgreiche magnetische und fluoreszierende DoppelmarkieAbstract ! Purpose: The aim of this study was to establish co-labeling of mesenchymal stromal cells (MSC) for the detection of single MSC in-vivo by MRI and histological validation. Materials and Methods: Mouse MSC were co-labeled with fluorescent iron oxide micro-particles and carboxyfluorescein succinimidyl ester (CFSE). The cellular iron content was determined by atomic absorption spectrometry. Cell proliferation and expression of characteristic surface markers were determined by flow cytometry. The chondrogenic differentiation capacity was assessed. Different amounts of cells (n1 = 5000, n2 = 15 000, n3 = 50 000) were injected into the left heart ventricle of 12 mice. The animals underwent sequential MRI on a clinical 3.0 T scanner (Intera, Philips Medical Systems, Best, The Netherlands). For histological validation cryosections were examined by fluorescent microscopy. Results: Magnetic and fluorescent labeling of MSC was established (mean cellular iron content 23.6 ± 3 pg). Flow cytometry showed similar cell proliferation and receptor expression of labeled and unlabeled MSC. Chondrogenic differentiation of labeled MSC was verified. After cell injection MRI revealed multiple signal voids in the brain and fewer signal voids...

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 95%

Magnetic Resonance Imaging of Single Co-Labeled Mesenchymal Stromal Cells after Intracardial Injection in Mice

Salamon

Wicklein

Didié

et al. 2014

Fortschr Röntgenstr

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“…However, combinations of new approaches using complementarity region (CDR3) size spectratyping and high-resolution magnetic imaging using T cells labeled with physiologically inert nanoparticles should enable studies of adoptively transferred T cells in the absence of genetic marking. 56,[59][60][61] Therapeutic acceleration of T-cell recovery after SCT A number of promising strategies involving adoptive cell therapy are being explored in the setting of SCT. T-cell subsets are perturbed for months to years after SCT, 62 and the T-cell repertoire as assessed by CDR3 spectratyping is constricted and unstable after SCT.…”

Section: Engraftmentmentioning

confidence: 99%

T-cell reconstitution and expansion after hematopoietic stem cell transplantation: ‘T’ it up!

Porter

June

2005

Bone Marrow Transplant

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Summary:Adoptive immunotherapy is the isolation and infusion of antigen-specific or nonspecific lymphocytes. Adoptive therapy with T cells may have a role in replacing, repairing, or enhancing immune function damaged by cytotoxic therapies, and rapid lymphocyte recovery may improve outcome after autologous and allogeneic stem cell transplantation (SCT). Recently, a plethora of information on the basic mechanisms of T-cell biology and regulation of cellular immune responses has emerged, permitting the development of new forms of adoptive cell therapy. Efficient ex vivo culture method for T-cell subsets affords the possibility of adoptive transfer of T cells engineered with enhanced capacity for central memory, effector cytotoxicity, Th1, Th2, veto cell, and T regulatory functions. Studies show that homeostatic Tcell proliferation is important for effective adoptive immunotherapy and pretreatment with chemotherapy may enhance the effects of infused T cells. Replicative senescence, in part due to telomere erosion, likely limits successful adoptive immunotherapy, though it may be possible to maintain T-cell pools by enforced expression of telomerase. Clinical trials now demonstrate that it is possible to enhance immune reconstitution after SCT with cytokines or infusions of ex vivo costimulated expanded T cells. These data all support the premise that adoptive therapy can accelerate reconstitution of cellular immunity with enhanced antitumor effects following SCT. Bone Marrow Transplantation (2005) 35, 935-942.

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“…Although they provide less contrast enhancement per particle compared with SSPIO and MPIO, large numbers of particles can be loaded into each cell [18,19]. As cationic surfaces have been shown to facilitate cellular internalization [20,21], USPIO are often modified with polycationic cell permeating peptides (CPP) such as HIV transactivator (TAT) [22] or protamine [23]. Other transfection techniques, sometimes in concert with CPPs, are also used [24,25].…”

Section: Introductionmentioning

confidence: 99%

Size, charge and concentration dependent uptake of iron oxide particles by non-phagocytic cells

2008

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A promising new direction for contrast-enhanced magnetic resonance (MR) imaging involves tracking the migration and biodistribution of superparamagnetic iron oxide (SPIO)-labeled cells in vivo. Despite the large number of cell labeling studies that have been performed with SPIO particles of differing size and surface charge, it remains unclear which SPIO configuration provides optimal contrast in non-phagocytic cells. This is largely because contradictory findings have stemmed from the variability and imprecise control over surface charge, the general need and complexity of transfection and/or targeting agents, and the limited number of particle configurations examined in any given study. In the present study, we systematically evaluated the cellular uptake of SPIO in non-phagocytic T cells over a continuum of particle sizes ranging from 33 nm to nearly 1.5 μm, with precisely controlled surface properties, and without the need for transfection agents. SPIO labeling of T cells was analyzed by flow cytometry and contrast enhancement was determined by relaxometry. SPIO uptake was dose dependent and exhibited sigmoidal charge dependence, which was shown to saturate at different levels of functionalization. Efficient labeling of cells was observed for particles up to 300nm, however micron-sized particle uptake was limited. Our results show that an unconventional highly cationic particle configuration at 107 nm maximized MR contrast of T cells, outperforming the widely utilized USPIO (<50 nm).

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Efficient magnetic cell labeling with protamine sulfate complexed to ferumoxides for cellular MRI

Cited by 515 publications

References 26 publications

Magnetic Resonance Imaging of Single Co-Labeled Mesenchymal Stromal Cells after Intracardial Injection in Mice

Magnetic Resonance Imaging of Single Co-Labeled Mesenchymal Stromal Cells after Intracardial Injection in Mice

T-cell reconstitution and expansion after hematopoietic stem cell transplantation: ‘T’ it up!

Size, charge and concentration dependent uptake of iron oxide particles by non-phagocytic cells

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