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Journal of Indian Association of Pediatric Surgeons
     Journal of Indian Association of Pediatric Surgeons
Official journal of the Indian Association of Pediatric Surgeons         
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Year : 2002  |  Volume : 7  |  Issue : 1  |  Page : 8-14

Hepatocyte transplantation in syngenic rat spleen


Dept. of Pediatric Surgery, All Institute of Medical Sciences, New Delhi-110029, India

Correspondence Address:
S Dave
Dept. of Pediatric Surgery, All Institute of Medical Sciences, New Delhi-110029
India
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Source of Support: None, Conflict of Interest: None


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ABSTRACT: Hepatocyte transplantation (HT) has generated interest because of potential clinical applications especially in fulminant hepatic failure and in ex-vivo gene therapy. Studies conducted in recent years have shown survival of heterotopically transplanted hepatocytes with improved survival in acute hepatic failure and correction of congenital enzyme deficiency diseases. The present study aims to establish a murine model for intrasplenic hepatocyte transplantation and confirm long term survival of hepatocytes by histopathological examination and by in vivo 99m Tc-HIDA scintigraphy. The study used inbred Fisher rats for hepatocyte isolation and transplantation. Hepatocyte isolation was carried out using a modified collagenase digestion method to achieve a 85 cell viability. Hepatocyte transplantation was performed in 6 rats by injecting 0.5ml (8 X 10 to the power 6 cells ) of the isolated hepatocyte suspension into the splenic parenchyma. The control group of 4 rats were injected with 0.5 ml normal saline. In vivo dynamic 99m Tc-HIDA scintigraphy was performed 90 days after transplantation. The rats were sacrificed and the spleen was evaluated histopathologically using routine H & E staining and periodic acid Schiff (PAS) stain for glycogen deposits. The scintigraphic studies showed that 5 of the 6 transplanted rats demonstrated uptake of HIDA in contrast to none of the control animals. Further, the time activity curves of the liver and spleen of transplanted rats showed similar kinetic patterns. Histological evaluation showed hepatocytes in the splenic red pulp with fatty vacuolation and PAS positive glycogen granules in the transplanted group of rats. In conclusion, hepatocyte transplantation is feasible in syngenic rat spleens and the collagenase digestion method adopted in this study, achieves a high yield of viable cells. In vivo dynamic HIDA scintigraphy is an easy and reliable modality for noninvasive graft function monitoring.






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  2005 - Journal of Indian Association of Pediatric Surgeons | Published by Wolters Kluwer - Medknow 

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