Labeling of Primary Human Hepatocytes With Micron‐Sized Iron Oxide Particles in Suspension Culture Suitable for Large‐Scale Preparation

Abstract

Labeling of hepatocytes with micron-sized iron oxide particles (MPIOs) enables cell detection using clinical magnetic resonance equipment. For clinical applications, large numbers of cells must be labeled in a simple and rapid manner and have to be applied in suspension. However, all existing protocols are based on adhesion culture labeling with subsequent resuspension, only suitable for small experimental settings. The aim of this study was to investigate the feasibility of preparing MPIO-labeled primary human hepatocytes in a temporary suspension culture. Human hepatocytes were isolated from 16 donors and labeled with MPIOs in suspension, using the Rotary Cell Culture System. Particle incorporation was investigated by light and electron microscopy. Cells were compared with adhesion culture-labeled and subsequently enzymatically resuspended cells. During a period of 5 days, hepatocyte-specific parameters of cell damage (aspartate aminotransferase and alanine aminotransferase) and metabolic activity (urea and albumin) were analyzed (n=7). Suspension cultures showed a higher outcome in cell recovery compared with the conventional labeling method. When incubated with 180 particles/viable cell for 4 h, the mean particle uptake was 28.8 particles/cell at a labeling efficiency of 95.1%. Labeling in suspension had no adverse effects on cell integrity or metabolic activity. We conclude that labeling of human hepatocytes in suspension is feasible and simple and may serve future large-scale processing of cells.

Clinical human hepatocyte transplantation: Current status and challenges

“…Others have used larger micrometer-sized iron oxide particles, which are not yet available for clinical use but have the advantage that hepatocytes can be labeled in suspension. 39 When given to mice, the iron particles were detected in Kupffer cells after 7 days rather than in hepatocytes. 40 Further work is needed to translate this approach to hepatocyte labeling for clinical use.…”

“…Cell‐labeling experiments in rats with ALF were performed and are currently being analyzed. Others have used larger micrometer‐sized iron oxide particles, which are not yet available for clinical use but have the advantage that hepatocytes can be labeled in suspension . When given to mice, the iron particles were detected in Kupffer cells after 7 days rather than in hepatocytes .…”

Artificial Organs 2011: A Year in Review

“…Nora N. Kammer et al. (97) of the Universitätsmedizin Berlin, Leipzig, Germany investigated the feasibility of preparing micron‐sized iron oxide particles (MPIOs)‐labeled primary human hepatocytes for eventual use in clinical magnetic resonance. Human hepatocytes were isolated from 16 donors and labeled with MPIOs in suspension.…”