|Shapiro, Erik - Michigan State University|
Submitted to: PLoS One
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/9/2015
Publication Date: 4/9/2015
Citation: Roach, D.R., Garrett, W.M., Welch, G.R., Caperna, T.J., Talbot, N.C., Shapiro, E.M. 2015. Magentic Cell labeling of primary and stem cell-derived pig hepatocytes for MRI-based cell tracking of heptocytes transplantation. PLoS One. 10(4):e0123282. DOI: 10.1371/journal.pone.0123282. eCollection 2015.
Interpretive Summary: Liver cell transplantation is a promising regenerative medicine strategy to cure liver diseases. The goal of this study is to use the pig as a large animal model to investigate several aspects of liver cell transplantation. In addition to defining and characterizing the optimal cell type, paramount to achieving successful transplants is the ability to track individual cells once they have been injected into live subjects. In this study we have developed a strategy to efficiently label ARS PICM-19 pig liver stem cells and cells isolated from baby pig livers with magnetic particles, so that they can be tracked by magnetic resonance imaging or MRI. These particles were shown to be rapidly taken up by both cell types and did not alter cell morphology or metabolic processes. The results pave the way for the next phase of this research which involves injecting magnetic particle-labeled cells into normal pigs and tracking them by MRI for several days. Once the conditions for successful transplantation are established, cells will be injected into piglets with liver disease to determine whether the transplanted cells can restore normal liver function.
Technical Abstract: Pig hepatocytes are an important investigational tool for optimizing hepatocyte transplantation schemes in both allogeneic and xenogeneic transplant scenarios. MRI can be used to serially monitor the transplanted cells, but only if the hepatocytes can be labeled with a magnetic particle. In this work, we describe culture conditions for magnetic cell labeling of cells from two different pig hepatocyte cell sources; primary pig hepatocytes (ppHEP) and stem cell-derived hepatocytes (PICM-19FF). The magnetic particle is a micron-sized iron oxide particle, or MPIO, that has been extensively studied for magnetic cell labeling for MRI-based cell tracking. ppHEP could endocytose MPIO with labeling percentages as high as 70%, achieving iron content as high as ~55 pg/cell, with >75% viability. PICM-19FF had labeling >97%, achieving iron content ~38 pg/cell, with viability >99%. Extensive morphological and functional assays indicated that magnetic cell labeling was benign to the cells. CONCLUSIONS: Magnetic cell labeling of ppHEP and PICM-19FF cells is innocuous to phenotype and important cell functions. These results encourage the use of MRI-based cell tracking for the development and clinical use of hepatocyte transplantation methodologies.