In Vitro Detection of Residual Undifferentiated Cells in Retinal Pigment Epithelial Cells Derived from Human Induced Pluripotent Stem Cells
In Vitro Detection of Residual Undifferentiated Cells in Retinal Pigment Epithelial Cells Derived from Human Induced Pluripotent Stem Cells
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DOI：
10.1007/978-1-4939-1435-7
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Sato, Yoji
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文献类型：
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作者：
Kuroda, Takuya;Yasuda, Satoshi;Sato, Yoji
人类多能干细胞(HPSCs)如人胚胎干细胞(HESCs)和人诱导多能干细胞(HiPSCs)因其具有多能性和无限自我更新能力而成为再生医学/细胞治疗的主要候选细胞。然而，阻碍hPSCs临床应用的因素很多。一个重要的安全问题是存在残留的未分化细胞，这些细胞有可能在体内形成肿瘤。在这里，我们描述了高灵敏的定量逆转录聚合酶链式反应方法来检测视网膜色素上皮(RPE)细胞中残留的未分化细胞。使用针对Lin28a(LIN28)转录本的探针和引物的QRT-PCR法可以检测到HiPSC来源的视网膜色素上皮细胞中低至0.002%的未分化细胞残留水平。我们期望该方法有助于HiPSC来源的细胞治疗产品的工艺验证和质量控制。
Human pluripotent stem cells (hPSCs) such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) are a leading candidate for regenerative medicine/cell therapies because of their capacity for pluripotency and unlimited self-renewal. However, there are significant obstacles preventing the clinical use of hPSCs. A significant safety issues is the presence of residual undifferentiated cells that have the potential to form tumors in vivo. Here, we describe the highly sensitive qRT-PCR methods for detection of residual undifferentiated cells in retinal pigment epithelial (RPE) cells derived from hiPSCs. qRT-PCR using probes and primers targeting LIN28A (LIN28) transcripts can detect residual undifferentiated cell levels as low as 0.002 % in hiPSC-derived RPE cells. We expect this method to contribute to process validation and quality control of hiPSC-derived cell therapy product.