Minami Takahito, Ishii Takamichi, Yasuchika Kentaro, Fukumitsu Ken, Ogiso Satoshi, Miyauchi Yuya, Kojima Hidenobu, Kawai Takayuki, Yamaoka Ryoya, Oshima Yu, Kawamoto Hiroshi, Kotaka Maki, Yasuda Katsutaro, Osafune Kenji, Uemoto Shinji
Department of Surgery, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Regen Ther. 2019 Mar 30;10:127-133. doi: 10.1016/j.reth.2019.03.002. eCollection 2019 Jun.
Liver transplantation is currently the only curative therapy for end-stage liver failure; however, establishment of alternative treatments is required owing to the serious donor organ shortage. Here, we propose a novel model of hybrid three-dimensional artificial livers using both human induced pluripotent stem cells (hiPSCs) and a rat decellularized liver serving as a scaffold.
Rat liver harvesting and decellularization were performed as reported in our previous studies. The decellularized liver scaffold was recellularized with hiPSC-derived hepatocyte-like cells (hiPSC-HLCs) through the biliary duct. The recellularized liver graft was continuously perfused with the culture medium using a pump at a flow rate of 0.5 mL/min in a standard CO (5%) cell incubator at 37 °C.
After 48 h of continuous perfusion culture, the hiPSC-HLCs of the recellularized liver distributed into the parenchymal space. Furthermore, the recellularized liver expressed the albumin () and genes, and secreted human ALB into the culture medium.
Novel hybrid artificial livers using hiPSCs and rat decellularized liver scaffolds were successfully generated, which possessed human hepatic functions.
肝移植是目前治疗终末期肝衰竭的唯一治愈性疗法;然而,由于供体器官严重短缺,需要建立替代治疗方法。在此,我们提出一种新型的混合三维人工肝模型,该模型使用人类诱导多能干细胞(hiPSC)和大鼠去细胞肝脏作为支架。
大鼠肝脏的获取和去细胞处理按照我们之前研究中的报道进行。通过胆管将hiPSC来源的肝样细胞(hiPSC-HLC)重新接种到去细胞肝脏支架上。在37℃的标准CO₂(5%)细胞培养箱中,使用泵以0.5 mL/min的流速对重新接种细胞的肝脏移植物持续灌注培养基。
连续灌注培养48小时后,重新接种细胞的肝脏中的hiPSC-HLC分布到实质间隙中。此外,重新接种细胞的肝脏表达白蛋白()和基因,并将人ALB分泌到培养基中。
成功构建了使用hiPSC和大鼠去细胞肝脏支架的新型混合人工肝,其具有人类肝脏功能。