Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material’s cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7–13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.

中文翻译：

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发光氧指示剂的共价固定降低了细胞毒性。

基于发光的氧感测是细胞培养应用中广泛使用的工具。在典型配置中，发光氧指示剂嵌入与培养基接触的固体，可透氧的基质中。然而，在敏感的细胞培养物中，即使最小程度地浸出潜在的细胞毒性指示剂也可能成为问题。防止浸出的一种方法是将指示剂共价固定在支持基质中。在本文中，我们报告了一种将铂（II）-5,10,15,20-四-（2,3,4,5,6-五氟苯基）-卟啉（PtTFPP）氧指示剂共价固定在由聚苯乙烯和聚五氟苯乙烯组成的聚合物基体。我们通过7到13天的培养实验和各种生存力分析，研究共价固定化如何影响传感材料对人类诱导的多能干细胞衍生（hiPSC衍生）神经元和心肌细胞（CMs）的细胞毒性。此外，我们研究了共价固定化对指示剂浸出和材料的氧传感特性的影响。另外，我们证明了在hiPSC衍生的CM的功能性低氧研究中，实时氧张力监测中共价连接的氧传感材料的使用。结果表明，共价固定的指示剂大大减少了指示剂的浸出和氧传感材料的细胞毒性，而对氧传感性能的影响仍然很小或不存在。