Hans Schöler Stem Cell Research Center (HSSCRC), School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, South Korea.
Group of Computational Biology and Bioinformatics, Biodonostia Health Research Institute, San Sebastián, and IKERBASQUE, Basque Foundation for Science, Bilbao, Spain.
PLoS One. 2019 Aug 12;14(8):e0221085. doi: 10.1371/journal.pone.0221085. eCollection 2019.
Direct conversion from fibroblasts to generate hepatocyte like-cells (iHeps) bypassing the pluripotent state has been described in previous reports as an attractive method acquiring hepatocytes for cell-based therapy. The limited proliferation of iHeps, however, has hampered it uses in cell-based therapy. Since hepatic stem cells (HepSCs) possess self-renewal and bipotency with the capacity to differentiate into both hepatocytes and cholangiocytes, they have therapeutic potential for treating liver disease. Here, we investigated the therapeutic effects of induced HepSCs (iHepSCs) on a carbon tetrachloride (CCl4)-induced liver fibrosis model. We demonstrate that Oct4 and Hnf4a are sufficient to convert fibroblasts into expandable iHepSCs. Hepatocyte-like cells derived from iHepSCs (iHepSC-HEPs) exhibit the typical morphology of hepatocytes and hepatic functions, including glycogen storage, low-density lipoprotein (LDL) uptake, Indocyanine green (ICG) detoxification, drug metabolism, urea production, and albumin secretion. iHepSCs-derived cholangiocyte-like cells (iHepSC-CLCs) expressed cholangiocyte-specific markers and formed cysts and tubule-like structures with apical-basal polarity and secretory function in three-dimensional culture condition. Furthermore, iHepSCs showed anti-inflammatory and anti-fibrotic effects in CCl4-induced liver fibrosis. This study demonstrates that Oct4 and Hnf4α-induced HepSCs show typical hepatic and biliary functionality in vitro. It also presents the therapeutic effect of iHepSCs in liver fibrosis. Therefore, directly converting iHepSCs from somatic cells may facilitate the development of patient-specific cell-based therapy for chronic liver damage.
直接将成纤维细胞转化为肝样细胞（iHeps），从而绕过多能状态，在以前的研究中被描述为一种获取用于细胞治疗的肝细胞的有吸引力的方法。然而，iHeps 的有限增殖阻碍了其在细胞治疗中的应用。由于肝干细胞（HepSCs）具有自我更新和多能性，能够分化为肝细胞和胆管细胞，因此它们具有治疗肝脏疾病的潜力。在这里，我们研究了诱导的肝干细胞（iHepSCs）对四氯化碳（CCl4）诱导的肝纤维化模型的治疗效果。我们证明 Oct4 和 Hnf4a 足以将成纤维细胞转化为可扩增的 iHepSCs。iHepSCs 衍生的肝样细胞（iHepSC-HEPs）表现出典型的肝细胞形态和肝脏功能，包括糖原储存、低密度脂蛋白（LDL）摄取、吲哚菁绿（ICG）解毒、药物代谢、尿素生成和白蛋白分泌。iHepSCs 衍生的胆管样细胞（iHepSC-CLCs）表达胆管细胞特异性标志物，并在三维培养条件下形成具有顶底极性和分泌功能的胆管样细胞囊和管状结构。此外，iHepSCs 在 CCl4 诱导的肝纤维化中显示出抗炎和抗纤维化作用。本研究表明，Oct4 和 Hnf4α 诱导的 HepSCs 在体外具有典型的肝和胆管功能。它还展示了 iHepSCs 在肝纤维化中的治疗效果。因此，直接将体细胞转化为 iHepSCs 可能有助于开发针对慢性肝损伤的患者特异性细胞治疗。
