Document Type : Original Article
Authors
1
Departments of Biochemistry and Molecular Biology, Theodor Bilharz Research Institute, Academy of Scientific Research and Technology, Cairo, Egypt.
2
Department of Surgical Research and Transplantology Laboratories, Medical Research Center, Polish Academy of Sciences, Warsaw, Poland.
3
Clinical and Experimental Surgery, Theodor Bilharz Research Institute, Academy of Scientific Research and Technology, Cairo, Egypt.
4
Department of Pathology, Theodor Bilharz Research Institute, Academy of Scientific Research and Technology, Cairo, Egypt.
5
Department of Immunology, Theodor Bilharz Research Institute, Academy of Scientific Research and Technology, Cairo, Egypt.
Abstract
Orthotropic Liver transplantation (OLT) is a conventional management for end-stage acute or chronic liver insufficiency, but the shortage of donor organs continues to be the restrictive factor throughout the world. Hepatocyte transplantation (HCTx) might be the promising treatment for several liver diseases and can be used as a "bridge" to OLT. Hepatocytes transplantation can protect and even save human lives, its’ applicability remains limited by the large deficiency of liver organs and hepatocytes (HC), and cellular loss after engraftment. Host elimination of grafted cells is called Early Graft Dysfunction. This study was developed for an efficient protocol of HCT. Several conditions have been met in order to achieve a high yield of harvested viable HC, overcome the detachedcell apoptosis, attenuation of innate immune reaction against transplanted cells and a receptive cell environment. HC were isolated from Lewis rats (n=8) weighing 250gm, by
the 2 step collagen a seper fusion technique, and bone marrow cells (BMCs) were obtained from the rats tibia and femur by centrifugation in a buffer solution. The mean viability of harvested HC and BMCs were 90% and 95% respectively.
To minimize the rejection of HC, Lewis rat recipients (n=14) weighing 250gm, were irradiated with 6 Gy and received 0.1 mg of anti-aisle GM1 antiserum intravenously as immunosuppressive drug. The isolated HC were intra-splenically transplanted and 107 bone marrow cells were injected in a penile vein into the recipients on the third day. Simultaneously,
70% hepatectomy and ligation of common bile duct were done. Thirty days later; the grafted spleen had areas with external appearance of a normal liver in ten out of 14surviving rats (71%). Hematoxlin and eosin (H & E) staining of sections from these fragments showed sinusoids and portal areas, an evidence of successful hepatocyte engraftment
and bile canaliculea formation. Large number of HC clusters of 15 to 20 cells and 2 to 4 distended small bile canaliculea were seen per50 HC. The intrasplenic route for transplanting freshly isolated HC in an immune-compromised animal model was found to give good results regarding cell engraftment and tissue formation.
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