Department of histology, Faculty of medicine, Ain Shams University.
Abstract
Introduction: The need for liver transplantation is rising due to increase in end-stage liver diseases. Testing drug toxicity on either liver tissue or on adult hepatocytes is crucial as it is the major drug metabolizing organ. Aim of the Work: This study aimed to form natural liver scaffold by decellularization followed by recellularization using adult male albino rat hepatocytes to form auxiliary hepatic tissue, and to culture adult hepatocytes for experimental studies and liver bioengineering. Material and Methods: Group I: [10 rats] control group. Group II: [10 rats] liver decellularization group. Group III: [20 rats] 10 rats used as adult hepatocyte donor for recellularization, and some hepatocytes were cultivated in culture flasks to ensure proliferation, while another 10 rats’ livers were decellularized followed by liver recellularization. At the end of the experiment, livers of control, decellularized scaffold, and recellularized tissue were processed for histological, histochemical and immunohistochemical techniques. Morphometrical and statistical studies were also performed. Results: Grossly, decellularized livers appeared semi-transparent. Histologically they appeared as an acellular liver matrix with apparently preserved hepatic architecture. The cultivated hepatocytes in culture flasks showed progressively increased number till confluency at day 21. They appeared polygonal with granular cytoplasm when cocultured with other cells. Recellularized livers showed hepatocyte aggregations with satisfactory preservation of extracellular matrix. Reseeded hepatocytes appeared polygonal with acidophilic, granular and Periodic acid-Schiff positive cytoplasm. Also, they showed significant increase in mean number of cells exhibiting positive reaction for proliferating cell nuclear antigen and Alpha-fetoprotein immune-stained sections compared to control. Conclusion: Cultivated adult rat hepatocytes proliferated till confluency in culture flasks. Well-structured decellularized tissue was formed that supported hepatocyte survival and proliferation, thus, this recellularized liver model and the cultured hepatocytes can be used for pharmaceutical testing on liver models and could be a new hope for patients with liver diseases.
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