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Labeling Protocols for In Vivo Tracking of Human Skeletal Muscle Cells (HSkMCs) by Magnetic Resonance and Bioluminescence Imaging

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Abstract

Purpose

We propose herein labeling protocols for multimodal in vivo visualization of human skeletal muscle cells (HSkMCs) by MRI and BLI to investigate the survival, localization, and proliferation/differentiation of these cells in cell-mediated therapy.

Procedures

HSkMCs were labeled with different quantities of Endorem® and transfection agents or infected with lentiviral vector expressing the luciferase gene under the myogenin promoter. Cells were evaluated before and after intra-arterial injection in NUDE mice with N2-induced muscle inflammation.

Results

Neither iron labeling nor infection affected cell features; the number of iron-positive cells increased proportionally to the iron content in the medium and in the presence of transfection agents. Loaded cells were detected for up to 1 month by MRI and 2 months by BLI.

Conclusions

These protocols could be used to visualize new stem cells, in vivo and over time, in preclinical studies of cell-based treatments for myopathies of different etiologies.

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Abbreviations

FLASH 3D:

Fast low-angle shot 3-dimensional sequence

i.a.:

Intra-arterial

i.m.:

Intramuscular

HSkMCs:

Human skeletal muscle cells

MRI:

Magnetic resonance imaging

MSCs:

Mesenchymal stem cells

MSME:

Multi-slice multi-echo

NSCs:

Neuronal stem cells

N2 :

Liquid nitrogen

o.m.:

Original magnification

PB:

Hexadimethrine bromide (polybrene)

PLL:

Poly-l-lysine hydrobromide

PrS:

Protamine sulfate

SPIO:

Superparamagnetic iron oxide

T2 :

Time of loss of transverse magnetization

ESCs:

Embryonic stem cells

iPSCs:

Induced pluripotent stem cells

BMSCs:

Bone marrow-derived stem cells

ASCs:

Adult stem cells

USPIO:

Ultra-small superparamagnetic iron oxide

BLI:

Bioluminescence imaging

MuSCs:

Muscle stem cells

NTX:

Notexin

DAB:

Diaminobenzidine

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Acknowledgments

We wish to thank Dr. B. Rosa (Accelera S.r.l.) for surgical training, Dr. M. Saresella and Dr. R. Lui for their help with the FACS analysis, D. Tosi for the IHC images, Dr. F. Corsi and R. Allevi for the electron microscopy (Centre of Electron Microscopy for the Development of Medical Nanotechnologies, L. Sacco Hospital Via G.B. Grassi), and Dr. S. Rivella for providing the PLW lentiviral backbone. The authors are also grateful to Mrs. Catherine Wrenn for her advice and skillful editorial support.

This study was supported by a Cariplo Foundation grant (2007.5281)

The authors declare that they have no conflict of interest

Author information

Authors and Affiliations

  1. Department of Biomedical Sciences and Technologies, Section of Radiological Sciences, University of Milan, Via di Rudinì 8, 20142, Milan, Italy

    Ilaria V. Libani, Giovanni Lucignani & Luisa Ottobrini

  2. Centre of Molecular and Cellular Imaging—IMAGO, University of Milan, via Fratelli Cervi 93, 20090, Segrate, MI, Italy

    Giovanni Lucignani, Mario Clerici & Luisa Ottobrini

  3. Pathology Unit, Department of Medicine, Surgery and Dentistry, University of Milan Medical School, IRCCS Ca’ Granda—Ospedale Maggiore Policlinico Foundation, Milan, Italy

    Umberto Gianelli & Silvano Bosari

  4. Pharmacology Department, BU Oncology, Nerviano Medical Sciences, v.le Pasteur 10, 20014, Nerviano, MI, Italy

    Anna Degrassi & Micaela Russo

  5. Department of Biomedical Sciences and Technologies, University of Milan, via Fratelli Cervi 93, 20090, Segrate, MI, Italy

    Mario Clerici

  6. Don C. Gnocchi Foundation IRCCS, Milan, Via Capecelatro 66, 20148, Milan, Italy

    Mario Clerici

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  1. Ilaria V. Libani
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  2. Giovanni Lucignani
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  8. Luisa Ottobrini
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Correspondence to Mario Clerici.

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Libani, I.V., Lucignani, G., Gianelli, U. et al. Labeling Protocols for In Vivo Tracking of Human Skeletal Muscle Cells (HSkMCs) by Magnetic Resonance and Bioluminescence Imaging. Mol Imaging Biol 14, 47–59 (2012). https://doi.org/10.1007/s11307-011-0474-6

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  • DOI: https://doi.org/10.1007/s11307-011-0474-6

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