Hepatocyte labeling with Tc-99m-GSA: a potential non-invasive technique for tracking cell transplantation

作者：Chouhan Manil; Puppi Juliana; Solanas Estela; Mitry Ragai R; Dhawan Anil; Hughes Robin D^*

来源：The International Journal of Artificial Organs, 2012, 35(6): 450-457.

DOI：10.5301/ijao.5000096

• Transplantation
• Galactosyl serum albumin
• Hepatocytes
• Cell labeling
• Radioactive technetium
• Asialoglycoprotein receptor

Background: Hepatocyte transplantation is a promising alternative to orthotopic liver transplantation, however, the fate of transplanted hepatocytes is not well defined. Tc-99m-galactosyl-serum albumin (Tc-99m-GSA) is a clinical scintigraphic agent which is specifically taken up by the hepatocyte asialoglycoprotein receptor (ASGPR).
Aims: To investigate labeling of fresh and cryopreserved human hepatocytes and fresh rat hepatocytes in vitro using Tc-99m-GSA.
Methods: Human and rat hepatocytes.were isolated from liver tissue by collagenase perfusion. The ASGPR were characterized using immunohistochemistry and RT-PCR. Hepatocytes were incubated with Tc-99m-GSA in suspension at 4 degrees C and 37 degrees C. Cell viability and function was determined using cell mitochondrial dehydrogenase (MTS) and sulphorhodamine B (SRB) assays.
Results: Fresh and cryopreserved human hepatocytes expressed the ASGPR. Incubation of hepatocytes in suspension with Tc-99m-GSA reduced the viability of hepatocytes, but this was similar to unlabeled control cells. Greater loss of viability was seen on incubation at 37 degrees C compared to 4 degrees C, but there was a significantly greater uptake of Tc-99m-GSA at the physiological temperature (6.6 +/- SE 0.6-fold increase, p < 0.05) consistent with ASGPR-mediated endocytosis. MTS and SRB assays were not significantly affected by labeling with Tc-99m-GSA in all three cell types. A mean of 18.5% of the radioactivity was released over 120 min when Tc-99m-GSA -labeled hepatocytes were shaken in vitro at 37 degrees C.
Conclusions: Human and rat hepatocytes can be labeled with Tc-99m-GSA, which may have potential application for in vivo imaging after hepatocyte transplantation.

• 出版日期2012-6