Induction of Pluripotent Stem Cell-Derived Cardiomyocyte Toxicity by Supernatant of Long Term-Stored Red Blood Cells in Vitro
Induction of Pluripotent Stem Cell-Derived Cardiomyocyte Toxicity by Supernatant of Long Term-Stored Red Blood Cells in Vitro
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体外长期保存红细胞上清液诱导多能干细胞来源的心肌细胞毒性
DOI：
10.1159/000489073
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Wang, Deqing
中科院分区：
文献类型：
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作者：
Fan, Fengyan;Yu, Yang;Wang, Deqing
背景/目的：体外保存的红细胞（rbc）在储存过程中发生一系列形态、功能和代谢变化。红细胞代谢产物在储存过程中随时间积累，红细胞上清液（ssrbc）对组织细胞的毒性在很大程度上是未知的。在这里，我们旨在通过体外长期储存的红细胞上清研究心肌细胞毒性，并发现参与机制的元素。方法：采用人诱导多能干细胞衍生心肌细胞（hiPS-CMs）和实时细胞分析（RTCA）技术，分析d0、d14和d35 ss红细胞的心脏毒性。为了分析钾(K)和乳酸（LA）对ssrbc的心脏毒性，我们分别制备了相同浓度的钾和乳酸溶液，并与hiPS-CMs共培养。采用免疫荧光和hiPS-CMs基因表达阵列技术评价d35 K和d35 ss红细胞的作用。结果：d14、d35、d35 K溶液均可使hiPS-CM停止跳动。d14 SSRBC或d35 K存在，但d35 SSRBC不存在，48小时内心跳恢复；d0、d14和d35 LA溶液对其跳动模式无影响。处理48h后，免疫荧光结果显示纤维和肌节的完整性完好。基因表达阵列结果发现14个差异表达基因可能在细胞毒作用中发挥重要作用。结论：我们的研究结果表明，长期储存的ss红细胞在体外具有心肌细胞毒性。除了高钾诱导的心脏毒性外，长期储存的ss红细胞中一定还有其他未知成分对hiPS-CMs具有细胞毒性。(C) 2018作者：s . Karger AG，巴塞尔出版
Background/Aims: Preserved red blood cells (RBCs) in vitro undergo a series of morphological, functional and metabolic changes during storage. RBC metabolites accumulate over time during storage, the toxicity of the supernatants of RBCs (SSRBCs) on tissue cells is largely unknown. Here, we aimed to study cardiomyocyte toxicity by supernatant of long term-stored RBCs in vitro and to discover elements involved in the mechanism. Methods: Using human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) and real-time cell analyzing (RTCA), we analyzed the cardiotoxicity of d0, d14 and d35 SSRBCs. To analyze the cardiotoxicity of potassium (K) and lactic acid (LA) in SSRBCs, solutions containing the same concentrations of K and LA were respectively prepared and co-cultured with hiPS-CMs. Immunofluorescence and Gene Expression Array of hiPS-CMs were performed to evaluate the effects of d35 K and d35 SSRBCs. Results: The beating of hiPS-CM was stopped by d14, d35 SSRBCs, or d35 K solution. Beating resumed within 48 hours in the presence of d14 SSRBC or d35 K but not d35 SSRBC; d0, d14 and d35 LA solution had no effect on beating patterns. At 48h after treatment, the immunofluorescence results showed that the integrity of the filament and sarcomere were intact. Gene Expression Array results found 14 differentially expressed genes which were likely to play an important role in the cytotoxic effect. Conclusion: Our results demonstrated cardiomyocyte toxicity by long term-stored SSRBCs in vitro. Besides high K-induced cardiotoxicity, there must be other unknown components in long term-stored SSRBCs that are cytotoxic to hiPS-CMs. (C) 2018 The Author(s) Published by S. Karger AG, Basel