Stanford Cardiovascular Institute.
Department of Medicine, Division of Cardiology, and.
JCI Insight. 2021 Apr 8;6(7):142000. doi: 10.1172/jci.insight.142000.
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.
人类多能干细胞（PSCs）由胚胎干细胞（ESCs）和诱导多能干细胞（iPSCs）组成，为基于心脏细胞治疗的临床试验提供了机会。然而，一个必须克服的重要障碍是，由于分化批次中残留未分化 PSCs 的增殖能力，细胞移植后形成畸胎瘤的风险。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纯化剂，选择性地针对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，我们确定了一个合适的体外阿霉素剂量，（a）在细胞注射前消除残留的未分化干细胞，以防止细胞移植后形成畸胎瘤，（b）通过收缩性分析、电生理学、拓扑异构酶活性测定和活性氧生成的定量来证明不会导致 ESC 衍生的心肌细胞（CMs）的心脏毒性。这项研究建立了一种潜在的新方法，用于无致瘤性细胞治疗研究，旨在实现心脏细胞移植的临床应用。
