Department of Anatomy & Embryology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, the Netherlands.
Department of Molecular Epidemiology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, the Netherlands.
Stem Cell Reports. 2017 May 9;8(5):1340-1353. doi: 10.1016/j.stemcr.2017.03.009.
The ability to form teratomas in vivo containing multiple somatic cell types is regarded as functional evidence of pluripotency for human pluripotent stem cells (hPSCs). Since the Teratoma assay is animal dependent, laborious, and only qualitative, the PluriTest and the hPSC ScoreCard assay have been developed as in vitro alternatives. Here we compared normal hPSCs, induced hPSCs (hiPSCs) with reactivated reprogramming transgenes, and human embryonal carcinoma cells (hECs) in these assays. While normal hPSCs gave rise to typical teratomas, the xenografts of the hECs and the hiPSCs with reactivated reprogramming transgenes were largely undifferentiated and malignant. The hPSC ScoreCard assay confirmed the line-specific differentiation propensities in vitro. However, when undifferentiated cells were analyzed by the PluriTest, only hECs were identified as abnormal whereas all other cell lines were indistinguishable and resembled normal hPSCs. Our results indicate that pluripotency assays are best selected on the basis of intended downstream applications.
能够在体内形成包含多种体细胞类型的畸胎瘤被认为是人类多能干细胞（hPSC）多能性的功能证据。由于畸胎瘤检测依赖于动物，既费力又只能定性，因此开发了 PluriTest 和 hPSC 评分卡检测作为体外替代方法。在这里，我们比较了正常 hPSC、具有重新激活的重编程转基因的诱导 hPSC（hiPSC）和人胚胎癌细胞（hEC）在这些检测中的情况。虽然正常 hPSC 会产生典型的畸胎瘤，但 hEC 和具有重新激活的重编程转基因的 hiPSC 的异种移植物主要是未分化的和恶性的。hPSC 评分卡检测在体外证实了特定系的分化倾向。然而，当通过 PluriTest 分析未分化细胞时，只有 hEC 被鉴定为异常，而其他所有细胞系都无法区分，类似于正常 hPSC。我们的结果表明，多能性检测最好根据预期的下游应用进行选择。
