Detailed Characterization of Human Induced Pluripotent Stem Cells Manufactured for Therapeutic Applications
Detailed Characterization of Human Induced Pluripotent Stem Cells Manufactured for Therapeutic Applications
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DOI：
10.1007/s12015-016-9662-8
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发表时间：
2016-08-01
影响因子：
4.8
通讯作者：
Rao, Mahendra S.
中科院分区：
文献类型：
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作者：
Baghbaderani, Behnam Ahmadian;Syama, Adhikarla;Rao, Mahendra S.
我们最近已经描述了使用脐带血作为起始材料通过临床合规过程产生的人诱导多能干细胞（iPSC）主细胞库（MCB）的制造（Baghbaderani等人，Stem Cell Reports，5（4），647-659，2015）。在这篇手稿中，我们描述了使用该过程产生的两个iPSC克隆的详细表征，包括全基因组测序（WGS），微阵列和比较基因组杂交（aCGH）单核苷酸多态性（SNP）分析。我们比较了他们的配置文件与建议的校准材料，并与记者亚克隆和线从不同的捐助者通过类似的过程。我们相信，iPSC很可能被用于制造多种临床产品。我们进一步相信，作为输入材料的线条将在不同的地点使用，鉴于它们的不朽地位，将使用多年甚至数十年。因此，重要的是开发检测方法来监测细胞的状态及其在培养物中的漂移。我们建议，细胞的初始状态的详细表征，与一些校准材料的比较和报告sublcones的发展将有助于确定哪一组测试将是最有用的监测细胞和建立标准丢弃线。
We have recently described manufacturing of human induced pluripotent stem cells (iPSC) master cell banks (MCB) generated by a clinically compliant process using cord blood as a starting material (Baghbaderani et al. in Stem Cell Reports, 5(4), 647-659, 2015). In this manuscript, we describe the detailed characterization of the two iPSC clones generated using this process, including whole genome sequencing (WGS), microarray, and comparative genomic hybridization (aCGH) single nucleotide polymorphism (SNP) analysis. We compare their profiles with a proposed calibration material and with a reporter subclone and lines made by a similar process from different donors. We believe that iPSCs are likely to be used to make multiple clinical products. We further believe that the lines used as input material will be used at different sites and, given their immortal status, will be used for many years or even decades. Therefore, it will be important to develop assays to monitor the state of the cells and their drift in culture. We suggest that a detailed characterization of the initial status of the cells, a comparison with some calibration material and the development of reporter sublcones will help determine which set of tests will be most useful in monitoring the cells and establishing criteria for discarding a line.