Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.
Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Laboratory of Advanced Research D # 326, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.
Hum Cell. 2019 Apr;32(2):125-140. doi: 10.1007/s13577-018-00234-0. Epub 2019 Jan 12.
Liver transplantation is the most effective treatment for treating liver cirrhosis. However, a limited number of donors, graft rejection, and other complications can undermine transplant success. It is considered that cell transplantation is an alternative approach of liver transplantation. We previously developed a protocol for hepatic differentiation of cluster of differentiation 117+ stem cells isolated from human exfoliated deciduous tooth pulp (SHEDs) under hydrogen sulfide exposure. These cells showed excellent hepatic function. Here, we investigated whether hepatocyte-like cell transplantation is effective for treating carbon tetrachloride (CCl)-induced liver cirrhosis. SHEDs were hepatically differentiated, which was confirmed via immunological analyses and albumin concentration determination in the medium. Rats were intraperitoneally injected with CCl for and the differentiated cells were injected into rat spleen. Histopathological and immunohistochemical analyses were performed. Liver functions were serologically and pathologically determined. Quantitative real-time-polymerase chain reaction was implemented to clarify the treatment procedure of liver cirrhosis. In vitro-differentiated hepatocyte-like cells were positive for all examined hepatic markers. SHED-derived hepatocyte transplantation eliminated liver fibrosis and restored liver structure in rats. Liver immunohistochemical analyses showed the presence of human-specific hepatic markers, i.e., a large amount of human hepatic cells were very active in the liver and spleen. Serological tests revealed significant liver function recovery in the transplantation group. Expression of genes promoting fibrosis increased after cirrhosis induction but was suppressed after transplantation. Our results suggest that xenotransplantation of hepatocyte-like cells of human origin can treat cirrhosis. Moreover, cell-based therapy of chronic liver conditions may be an effective option.
肝移植是治疗肝硬化最有效的方法。然而，供体数量有限、移植物排斥和其他并发症可能会影响移植的成功。人们认为细胞移植是肝移植的一种替代方法。我们之前开发了一种在硫化氢暴露下从人脱落乳牙牙髓中分离的 CD117+干细胞分化为肝的方案。这些细胞表现出良好的肝功能。在这里，我们研究了肝细胞样细胞移植是否对治疗四氯化碳（CCl）诱导的肝纤维化有效。通过免疫分析和培养基中白蛋白浓度的测定，证实了 SHED 的肝分化。将 CCl 腹腔注射到大鼠体内，并将分化的细胞注射到大鼠脾脏中。进行了组织病理学和免疫组织化学分析。通过血清学和病理学方法确定肝功能。实施定量实时聚合酶链反应以阐明治疗肝纤维化的程序。体外分化的肝细胞样细胞对所有检查的肝标志物均呈阳性。SHED 来源的肝细胞移植消除了大鼠的肝纤维化并恢复了肝结构。肝免疫组织化学分析显示存在人类特异性肝标志物，即大量人类肝细胞在肝脏和脾脏中非常活跃。血清学检测显示移植组的肝功能明显恢复。诱导肝硬化后促进纤维化的基因表达增加，但移植后被抑制。我们的结果表明，源自人类的肝细胞样细胞的异种移植可以治疗肝硬化。此外，慢性肝脏疾病的细胞治疗可能是一种有效选择。
