Immunology Department, Theodor Bilharz Research Institute, Cairo, Egypt.
Pathology Department, and Theodor Bilharz Research Institute, Cairo, Egypt.
Cell Reprogram. 2020 Jun;22(3):134-146. doi: 10.1089/cell.2019.0076. Epub 2020 Apr 3.
Mesenchymal stem cell (MSC)-based liver tissue engineering on nanofibrous scaffold holds great promise for cell-based therapy in liver injuries and end-stage liver failure treatments. MSCs were generated from umbilical cord blood. Hepatogenic differentiation was induced on two-dimensional (2D) and three-dimensional (3D) culture system and characterized by morphology, scanning electron microscopy, immunocytochemistry, and gene expression. Albumin and α-1 antitrypsin (AAT) in culture supernatants were measured. Differentiated cells were administered intravenous into a murine model of carbon tetra induced liver cirrhosis. After 12 weeks of injection, liver pathology was examined. The hepatogenic differentiated MSCs stained positively for albumin, alpha fetoprotein, HepPar1, cytokeratin 7 and 18, and OV6 with more mature cells, hexagonal in shape with central nuclei forming large sheets in groups in 3D culture system. AAT secretion and indocyanine green uptake were significantly increased in 3D system. In experimental model, MSC-3D treated group exhibited maximal restoration of liver architecture with absent septal fibrosis and marked improvement of alanine transaminase (ALT) and aspartate transaminase (AST), and mild increase in albumin. Both 3D and 2D culture system are effective in functional hepatogenic differentiation from MSCs and serve as a vehicle in liver tissue engineering. hepatogenic differentiation is more effective on 3D scaffold, with better functional recovery.
基于间充质干细胞（MSC）的纳米纤维支架肝组织工程在细胞治疗肝损伤和终末期肝功能衰竭方面具有广阔的应用前景。MSC 是从脐带血中生成的。在二维（2D）和三维（3D）培养系统中诱导其向肝系细胞分化，并通过形态学、扫描电子显微镜、免疫细胞化学和基因表达进行鉴定。检测培养上清液中的白蛋白和α-1 抗胰蛋白酶（AAT）。将分化后的细胞经静脉注射到四氯化碳诱导的肝纤维化小鼠模型中。注射 12 周后，检查肝组织病理学。肝向分化的 MSC 在 3D 培养系统中呈阳性染色，表达白蛋白、甲胎蛋白、HepPar1、细胞角蛋白 7 和 18 以及 OV6，这些细胞更成熟，呈六边形，中央有核，在 3D 培养系统中成组形成大片。3D 系统中 AAT 分泌和吲哚菁绿摄取显著增加。在实验模型中，MSC-3D 治疗组表现出肝组织结构的最大恢复，不存在间隔纤维化，丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）显著改善，白蛋白轻度增加。3D 和 2D 培养系统都能有效地从 MSC 中进行功能性肝向分化，并作为肝组织工程的载体。3D 支架上的肝向分化更有效，功能恢复更好。
