In Vivo Fluorescence Visualization of Anterior Chamber Injected Human Corneal Endothelial Cells Labeled With Quantum Dots
In Vivo Fluorescence Visualization of Anterior Chamber Injected Human Corneal Endothelial Cells Labeled With Quantum Dots
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前房注射量子点标记的人角膜内皮细胞的体内荧光可视化
DOI：
10.1167/iovs.19-27788
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Hamuro Junji
中科院分区：
文献类型：
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作者：
Toda Munetoyo;Yukawa Hiroshi;Yamada Jun;Ueno Morio;Kinoshita Shigeru;Baba Yoshinobu;Hamuro Junji
目的：将培养的人角膜内皮细胞（cHCECs）注射到前房（AC）中是一种成功治疗角膜内皮功能障碍的新方法。在这里，我们研究了是否可以使用由半导体纳米颗粒八精氨酸（R8）组成的量子点（QD）标记cHCEC以追踪注射的cHCEC，并检查了体内荧光成像的实用性以分析QD标记的注射的cHCEC在角膜内皮功能障碍小鼠模型中的动力学和积累。通过添加QD 655和R8的混合物来标记高质量或具有细胞状态转变的cHCEC。通过流式细胞术证实了标记效率和通过标记的细胞表型的不变。将标记的cHCECs注射到健康小鼠或因冷冻处理而角膜内皮受损的小鼠的AC中。结果：QD标记后，cHCECs的形态及功能标志物Na+/K+-ATP酶和闭锁小带1的表达均无明显变化。定量检测到注射的cHCECs-QD，并且cHCECs-QD的保留是明显的，从细胞注射后3至48小时在低温损伤的角膜内皮小鼠模型眼中的后表面上，而在未损伤的健康对照眼中则没有。这项研究的结果表明，在再生医学领域，QD标记的细胞提供了一种方便和灵敏的方法，精细监测体内注射细胞的命运。
Purpose: The injection of cultured human corneal endothelial cells (cHCECs) into the anterior chamber (AC) is a newly developed modality for the successful treatment of corneal endothelium dysfunction. Here, we investigated whether or not cHCECs could be labeled using quantum dots (QDs) composed of semiconductor nanoparticle octa-arginine (R8) to trace injected cHCECs and examined the utility of in vivo fluorescence imaging to analyze the dynamics and accumulation of QD-labeled injected cHCECs in a corneal endothelial dysfunction mouse model.Methods: The cHCECs, either of high quality or with cell-state transition, were labeled by adding a mixture of QDs655 and R8. The labeling efficiency and the unchanging of the cell phenotypes by the labeling was confirmed by flow cytometry. The labeled cHCECs were injected into the AC of either healthy mice or mice with corneal endothelium damaged by cryogenic treatment. The kinetics of the injected cHCECs was traced quantitatively via multiphoton confocal laser microscopy.Results: QD labeling induced no morphologic change in the cHCECs or in the expression of the functional markers of cHCECs (ie, Na+/K+-ATPase and zonula occludens-1). The injected cHCECs-QDs were quantitatively detected, and the retention of cHCECs-QDs was evident, from 3 to 48 hours post cell injection on the posterior surface in the cryogenically injured corneal endothelium mouse model eyes, yet not in the noninjured healthy control eyes.Conclusions: The findings of this study show that in the field of regenerative medicine, QD labeling of cells presents a convenient and sensitive method of finely monitoring the fate of injected cells in vivo.