Method for selective ablation of undifferentiated human pluripotent stem cell populations for cell-based therapies
Method for selective ablation of undifferentiated human pluripotent stem cell populations for cell-based therapies
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DOI：
10.1172/jci.insight.142000
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发表时间：
2021-04-08
期刊：
影响因子：
8
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
--
作者：
Chour, Tony;Tian, Lei;Wu, Joseph C.
人类多能干细胞（PSC）由胚胎干细胞（ESC）和诱导多能干细胞（iPSC）组成，为推进基于心脏细胞治疗的临床试验提供了机会。然而，必须克服的一个重要障碍是细胞移植后由于分化批次中残留未分化PSC的增殖能力而形成畸胎瘤的风险。为了解决这个问题，我们建议使用最小的非心脏毒性阿霉素剂量作为净化剂，选择性地靶向快速增殖的干细胞进行细胞死亡，这将在细胞移植前提供更纯的终末分化心肌细胞群体。在这项研究中，我们确定了一个适当的体外阿霉素剂量，（a）消除残留的未分化的干细胞注射前，以防止畸胎瘤形成后，细胞移植和（B）不会导致心脏毒性的ESC衍生的心肌细胞（CM），通过收缩力分析，电生理学，拓扑异构酶活性测定，和定量的活性氧产生。本研究建立了一个潜在的新方法，无致瘤性细胞治疗的研究，旨在心脏细胞移植的临床应用。
Human pluripotent stem cells (PSCs), which are composed of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide an opportunity to advance cardiac cell therapy-based clinical trials. However, an important hurdle that must be overcome is the risk of teratoma formation after cell transplantation due to the proliferative capacity of residual undifferentiated PSCs in differentiation batches. To tackle this problem, we propose the use of a minimal noncardiotoxic doxorubicin dose as a purifying agent to selectively target rapidly proliferating stem cells for cell death, which will provide a purer population of terminally differentiated cardiomyocytes before cell transplantation. In this study, we determined an appropriate in vitro doxorubicin dose that (a) eliminates residual undifferentiated stem cells before cell injection to prevent teratoma formation after cell transplantation and (b) does not cause cardiotoxicity in ESC-derived cardiomyocytes (CMs) as demonstrated through contractility analysis, electrophysiology, topoisomerase activity assay, and quantification of reactive oxygen species generation. This study establishes a potentially novel method for tumorigenic-free cell therapy studies aimed at clinical applications of cardiac cell transplantation.