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

Chouhan, M. ; Puppi, J. ; Solanas, E. ; Mitry, R. ; Dhawan, A. ; Hughes, R.D.

Resumen: Background: Hepatocyte transplantation is a promising alternative to orthotopic liver transplantation, however, the fate of transplanted hepatocytes is not well defined. 99mTc-galactosyl-serum albumin (99mTc-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 99mTc-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 99mTc-GSA in suspension at 4°C and 37°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 99mTc-GSA reduced the viability of hepatocytes, but this was similar to unlabeled control cells. Greater loss of viability was seen on incubation at 37°C compared to 4°C, but there was a significantly greater uptake of 99mTc-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 99mTc-GSA in all three cell types. A mean of 18.5% of the radioactivity was released over 120 min when 99mTc-GSA - labeled hepatocytes were shaken in vitro at 37°C. Conclusions: Human and rat hepatocytes can be labeled with 99mTc-GSA, which may have potential application for in vivo imaging after hepatocyte transplantation.

Idioma: Inglés

DOI: 10.5301/ijao.5000096

Año: 2012

Publicado en: INTERNATIONAL JOURNAL OF ARTIFICIAL ORGANS 35, 6 (2012), 450-457
ISSN: 0391-3988

Factor impacto JCR: 1.759 (2012)

Categ. JCR: ENGINEERING, BIOMEDICAL rank: 35 / 76 = 0.461 (2012) - Q2 - T2
Categ. JCR: TRANSPLANTATION rank: 16 / 26 = 0.615 (2012) - Q3 - T2

Tipo y forma: Artículo (PostPrint)