Critical Functionality Effects from Storage Temperature on Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Suspensions
Critical Functionality Effects from Storage Temperature on Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Suspensions
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DOI：
10.1038/s41598-018-38065-6
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发表时间：
2019-02-27
影响因子：
4.6
通讯作者：
Takahashi, Masayo
中科院分区：
文献类型：
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作者：
Kitahata, Shohei;Tanaka, Yuji;Takahashi, Masayo
人诱导多能干细胞（hiPSC）来源的视网膜色素上皮（hiPSC- rpe）细胞悬浮液具有再生治疗的潜力。然而，hiPSC-RPE细胞的实际再生应用需要开发简单且具有成本效益的非冷冻保存方法。我们研究了不同条件下非冷冻温度对悬浮hiPSC-RPE细胞的影响，并分析了细胞死亡、凋亡、Rho gtpase、缺氧、微管破坏和细胞代谢的机制。保存在37℃的细胞由于高细胞代谢和细胞沉积的缺氧而具有最低的活力，保存在4℃的细胞由于微管脆性而受损。16℃时细胞悬液效果最佳，细胞凋亡显著减少，坏死可忽略不计。此外，与未保存的细胞相比，存活的细胞增殖和分泌关键蛋白质正常。hiPSC-RPE细胞悬液最佳保存温度为16℃，高于或低于最佳温度会显著降低细胞活力，但细胞死亡、细胞代谢、微管破坏和氧张力的机制不同，所有这些都与细胞状况有关。在高细胞死亡经常被报道的地方，存活的细胞有望作为移植物发挥作用。本研究为研究各种非冷冻温度对hiPSC-RPE细胞的影响提供了新的见解，这些影响与临床应用高度相关，并可能改善实验室和医院之间的合作。
Human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (hiPSC-RPE) cells suspension have the potential for regenerative treatment. However, practical regenerative applications with hiPSC-RPE cells require the development of simple and cost-effective non-freezing preservation methods. We investigated the effect of non-freezing temperatures on suspended hiPSC-RPE cells in various conditions and analysed mechanisms of cell death, anoikis, Rho GTPases, hypoxia, microtubule destruction, and cell metabolism. Cells stored at 37 degrees C had the lowest viability due to hypoxia from high cell metabolism and cell deposits, and cells preserved at 4 degrees C were damaged via microtubule fragility. Cell suspensions at 16 degrees C were optimal with drastically reduced apoptosis and negligible necrosis. Moreover, surviving cells proliferated and secreted key proteins normally, compared to cells without preservation. hiPSC-RPE cell suspensions were optimally preserved at 16 degrees C. Temperatures above or below the optimal temperature decreased cell viability significantly yet differentially by mechanisms of cell death, cellular metabolism, microtubule destruction, and oxygen tension, all relevant to cell conditions. Surviving cells are expected to function as grafts where high cell death is often reported. This study provides new insight into various non-freezing temperature effects on hiPSC-RPE cells that are highly relevant to clinical applications and may improve cooperation between laboratories and hospitals.