PREPARATION OF IRON OXIDE LABELLED PRIMARY HUMAN HEPATOCYTES SUITABLE FOR CELL TRANSPLANTATION

Aim: Transplantation of primary human hepatocytes is a promising approach for treatment of certain liver diseases. For a better understanding of the processes during and following transplantation, visualisation of the transplanted cells is desirable. Aim was the preparation of iron oxide labelled primary human hepatocytes for cell transplantation and the evaluation of this treatment on viability and metabolic activity of the cells. Methods: Primary human hepatocytes were isolated from liver specimen obtained from patients undergoing partial hepatectomy. Cells were incubated with fluorescence-labelled, micron-sized iron oxide particles (MPIO) at a concentration of 30×106 particles per million cells for 4 hours and re-suspended enzymatically. Incorporation of particles was assessed by light-, fluorescence- and electron microscopy and the cells were visualised using 3.0 Tesla MR scanner. Cells were re-cultivated and particle retention, viability and metabolic activity (AST, LDH, Urea, Albumin) were determined over a period of five days, thereafter. Results: The mean uptake was 18 particles per cell with 97% labelling efficiency. The particles were entirely incorporated into the cells and no evidence for particle release could be observed. Labelled cells were clearly detectable by T2*-weighted MR measurement. Labelling had no adverse effect on the viability or metabolic activity of primary human hepatocytes. Resuspended cells were cultivated successfully following the labelling procedure. Conclusion: This study presents the first protocol for preparation of iron oxide labelled primary human hepatocytes suitable for cell transplantation. Tracking and detection of primary human hepatocytes using MRI is an important step to improve the clinical application and describes a valuable tool for its quality control.