Amal Elham Fares1, Hala Gabr2, Asmaa Mohammed ShamsEldeen3, Haithem A M Farghali4, Mazen Mohamed Salem Mohamed Rizk5, Bassem Essam Mahmoud6, Ahmed Bakr Ahmed Tammam7, Ayman Magdy Ahmed Mahmoud8, Alaa Abdulfattah Mahmoud Suliman9, Mohamed Abdelhamid Ali Ayyad10, Sahar Hassan Ahmed11, Rokia Mohamad Hassan1. 1. Histology Department, Faculty of Medicine, Cairo University, Giza, Egypt. 2. Clinical Pathology Department, Faculty of Medicine, Cairo University, Giza, Egypt. 3. Physiology Department, Faculty of Medicine, Cairo University, Giza, Egypt. Asmaa82shamseldeen@gmail.com. 4. Surgery, Anesthesiology and Radiology Department, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt. 5. Faculty of Medicine, Cairo University, Giza, Egypt. 6. Siparadigm Diagnostics, Montville, NJ, USA. 7. Al Abbasya Hospital for Mental Health, Cairo, Egypt. 8. Anesthesia Department, Eldemerdash University Hospital, Cairo, Egypt. 9. Interventional and Diagnostic Radiology Department, Faculty of Medicine, Cairo University, Giza, Egypt. 10. National Cancer Institute, Cairo University, Giza, Egypt. 11. Medical Laboratory Technology Department, Faculty of Applied Health Science Technology, Misr University for Science and Technology, Giza, Egypt.
Abstract
BACKGROUND AND OBJECTIVES: The X-linked bleeding disorder, hemophilia A, is caused by defective production of factor VIII (FVIII). Hemophilic patients require regular FVIII infusions. Recombinant factor replacement poses the safest line of therapy. However, its main drawbacks are high expenses and the higher liability for formation of inhibitors. Recent studies confirmed the ability of bone marrow-derived stem cells to secrete FVIII. This study aims to generate bioscaffold from decellularized liver and subsequently seed it with trans-differentiated human stem cells into hepatic-like cells. This scaffold can then be implanted intraperitoneally or subcutaneously to provide FVIII. METHODS: After generation of the bioscaffold, seeding of discoid scaffolds with trans-differentiated human hepatocyte-like cells was performed. Then, the generated organoid was implanted into peritoneal cavity or subcutaneous tissue of experimental rats. RESULTS: Serum human FVIII was significantly increased in rats subjected to subcutaneous implantation compared intraperitoneal implantation. Immunostaining for detecting Cytokeratin 19 and human anti-globulin confirmed the presence of mature human hepatocytes that were significantly increased in subcutaneous implanted scaffold compared to the intraperitoneal one. CONCLUSION: Implantation of decellularized bioscaffold seeded with trans-differentiated stem cells in rats was successful to establish production of FVIII. Subcutaneous implantation showed higher FVIII levels than intraperitoneal implantation.
BACKGROUND AND OBJECTIVES: The X-linked bleeding disorder, hemophilia A, is caused by defective production of factor VIII (FVIII). Hemophilic patients require regular FVIII infusions. Recombinant factor replacement poses the safest line of therapy. However, its main drawbacks are high expenses and the higher liability for formation of inhibitors. Recent studies confirmed the ability of bone marrow-derived stem cells to secrete FVIII. This study aims to generate bioscaffold from decellularized liver and subsequently seed it with trans-differentiated human stem cells into hepatic-like cells. This scaffold can then be implanted intraperitoneally or subcutaneously to provide FVIII. METHODS: After generation of the bioscaffold, seeding of discoid scaffolds with trans-differentiated human hepatocyte-like cells was performed. Then, the generated organoid was implanted into peritoneal cavity or subcutaneous tissue of experimental rats. RESULTS: Serum humanFVIII was significantly increased in rats subjected to subcutaneous implantation compared intraperitoneal implantation. Immunostaining for detecting Cytokeratin 19 and human anti-globulin confirmed the presence of mature human hepatocytes that were significantly increased in subcutaneous implanted scaffold compared to the intraperitoneal one. CONCLUSION: Implantation of decellularized bioscaffold seeded with trans-differentiated stem cells in rats was successful to establish production of FVIII. Subcutaneous implantation showed higher FVIII levels than intraperitoneal implantation.