Abstract
Hemophilia is caused by a single-gene defect resulting in familial bleeding disorder. Small increase in gene products could transform a severe form of hemophilia into a mild one. Stem cells from extrahepatic sources are being considered for clinical applications in liver cell therapy as they possess high in vitro culture potential and could be used in transplant procedures. We studied the differentiation of bone marrow hemapoietic stem cells (BM-HSCs) from hemophilia patients' relatives into FVIII-producing hepatocytes aiming to expand patients' donor options for partial replacement of mutant liver cells by healthy cells in hemophilia A patients which could manage the severity of the bleeding disorder. BM-HSCs from hemophilic families were cultured in liquid culture containing hepatocyte growth factor for 6 days. Differentiation into hepatocytes was evaluated by alpha-fetoprotein (AFP) expression using immunocytochemistry. Functional evaluation of transdifferentiation into hepatic lineage was done through albumin synthesis in culture supernatant using microalbumin assay kit, factor VIII activity by one stage clotting assay and expression of FVIII mRNA by RT-PCR. BM-HSCs-derived hepatocytes showed positive AFP expression with a mean of 11 %. Functional tests performed showed their ability to produce albumin and perform FVIII activity. Also, FVIII mRNA expression was detected. Inducing the differentiation of BM-HSCs by in vitro manipulation may become a valuable tool to provide a cell source for hepatocyte transplant procedures for treatment of hemophilia patients.
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Gabr, H., Zayed, R., ElBeshlawy, A. et al. Differentiation of bone marrow hematopoietic stem cells into FVIII-producing hepatocytes: approach to hemophilia treatment. Comp Clin Pathol 23, 193–198 (2014). https://doi.org/10.1007/s00580-012-1595-2
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DOI: https://doi.org/10.1007/s00580-012-1595-2