Ichikawa Moe, Negoro Ryosuke, Kawai Kanae, Yamashita Tomoki, Takayama Kazuo, Mizuguchi Hiroyuki
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Laboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Mol Ther Methods Clin Dev. 2021 Jan 20;20:463-472. doi: 10.1016/j.omtm.2021.01.005. eCollection 2021 Mar 12.
Human induced pluripotent stem cell-derived intestinal epithelial cells (hiPSC-IECs) are expected to be utilized in regenerative medicine. To perform a safe transplantation without the risk of tumor formation, residual undifferentiated hiPSCs must be removed from hiPSC-IECs. In this study, we examined whether vinblastine (a multiple drug resistance 1 [MDR1] substrate) could remove residual undifferentiated hiPSCs in hiPSC-IECs and attempted to generate hiPSC-IECs applicable to transplantation medicine. We found that the expression levels of pluripotent markers were largely decreased and those of intestinal markers were increased by vinblastine treatment. The treatment of undifferentiated hiPSCs with vinblastine significantly decreased their viability. These results suggested that undifferentiated hiPSCs can be eliminated from hiPSC-IECs by vinblastine treatment. We hypothesized that MDR1-negative cells (such as undifferentiated hiPSCs) die upon vinblastine treatment because they are unable to excrete vinblastine. As expected, the cell viability of MDR1-knockout hiPSC-IECs was significantly decreased by vinblastine treatment. Furthermore, teratomas were formed by subcutaneous transplantation of hiPSC-IECs mixed with undifferentiated hiPSCs into mice, but they were not observed when the transplanted cells were pre-treated with vinblastine. Vinblastine-treated hiPSC-IECs would be an effective cell source for safe regenerative medicine.
人诱导多能干细胞来源的肠上皮细胞（hiPSC - IECs）有望用于再生医学。为了在无肿瘤形成风险的情况下进行安全移植，必须从hiPSC - IECs中去除残留的未分化hiPSC。在本研究中，我们检测了长春碱（一种多药耐药1 [MDR1]底物）是否能去除hiPSC - IECs中残留的未分化hiPSC，并试图生成适用于移植医学的hiPSC - IECs。我们发现，长春碱处理后多能性标志物的表达水平大幅下降，而肠道标志物的表达水平则升高。用长春碱处理未分化的hiPSC可显著降低其活力。这些结果表明，通过长春碱处理可从hiPSC - IECs中消除未分化的hiPSC。我们推测MDR1阴性细胞（如未分化的hiPSC）在长春碱处理后死亡是因为它们无法排出长春碱。正如预期的那样，长春碱处理使MDR1基因敲除的hiPSC - IECs的细胞活力显著下降。此外，将与未分化hiPSC混合的hiPSC - IECs皮下移植到小鼠体内会形成畸胎瘤，但在移植细胞用长春碱预处理时未观察到畸胎瘤。经长春碱处理的hiPSC - IECs将是安全再生医学的有效细胞来源。
