In Situ Labeling and Magnetic Resonance Imaging of Transplanted Human Hepatic Stem Cells
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The purpose is to address the problem in magnetic resonance imaging (MRI) of contrast agent dilution.
In situ magnetic labeling of cells and MRI were used to assess distribution and growth of human hepatic stem cells (hHpSCs) transplanted into severe combined immunodeficiency (SCID)/non-obese diabetic (NOD) mice. It was done with commercially available magnetic microbeads coupled to an antibody to a surface antigen, epithelial cell adhesion molecule (EpCAM), uniquely expressed in the liver by hepatic progenitors.
We validated the microbead connection to cells and related MRI data to optical microscopy observations in order to develop a means to quantitatively estimate cell numbers in the aggregates detected. Cell counts of hHpSCs at different times post-transplantation revealed quantifiable evidence of cell engraftment and expansion.
This magnetic labeling methodology can be used with any antibody coupled to a magnetic particle to target any surface antigen that distinguishes transplanted cells from host cells, thus facilitating studies that define methods and strategies for clinical cell therapy programs.
Key wordsMagnetic resonance imaging (MRI) Cell labeling and tracking Human hepatic stem cells Cell therapies EpCAM
Epithelial cell adhesion molecule
Hormonally defined medium
Human hepatic stem cell
Magnetically activated cell sorting
Magnetic resonance imaging
Technical and administrative support was provided by Lucendia English, Victoria Morgan, and Dr. Claire Barbier. The microscopy was done in the Michael Hooker Confocal Microscope Facility at UNC (Dr. Michael Chua, director) and the electron microscopy in the Microscope Facility (Dr. Robert Bagnell, director). We thank Dr. Sharon Lubkin for a critical evaluation of the paper and Dr. Claire Barbier for editing the figures.
This work was funded primarily by a US Department of Energy (DOE) grant (DE-FG02-02ER-63477). It derived also from grants from the North Carolina Biotechnology Center, Vesta Therapeutics (Bethesda, MD), the National Institutes of Health (NIH; AA014243 and IP30-DK065933), the National Institute of Diabetes and Digestive and Kidney Diseases (DK34987), and the National Cancer Institute (CA016086). All of the imaging was done in the Duke Center for In Vivo Microscopy (Dr. G.A. Johnson, director), an NIH/National Center for Research Resources Biomedical Technology Resource Center (P41 RR005959) and Small Animal Imaging Resource Program (U24 CA092656).
Conflict of Interest
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