Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging.
Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging.
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DOI：
10.1186/s13287-018-0909-y
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发表时间：
2018-06-13
影响因子：
7.5
通讯作者：
Paula JS
中科院分区：
文献类型：
--
作者：
Vilela CAP;Souza LEB;Siqueira RC;Calado RT;Covas DT;Paula JS
骨髓间充质基质细胞(MSC)治疗多种退行性眼病是一种很有前途的治疗方法；然而，目前还没有建立可重复使用的监测这些细胞进入眼内的方法。这项研究的目的是描述成功的生物发光成像(BLI)来检测眼睛中有活性的表达荧光素酶的MSC。眼内注射前，将50μL荧光素酶慢病毒载体(3个病毒颗粒/细胞)导入体外培养的人供体骨髓间充质干细胞。21只兔的21只右眼接受1 × _(106)荧光素酶表达的MSCs玻璃体腔内植入后行BLI检查。对侧眼注射磷酸盐缓冲液(PBS)作为对照。在7个不同的时间点(1小时至60天)，将d-荧光素(40 mg/ml，300μL缓冲液)注入6只摘除眼球的不同亚群，通过BLI分析评估辐射衰减。对选定的眼进行CD90和CD73免疫荧光检测。注射MSC的眼在注射d-荧光素后即刻表现为高BLI辐射，并逐渐腐烂，直至60天。从8h到30d，表达荧光素酶的MSC眼的平均BLI辐射测量显著高于对照组。注射MSC后第30天，仅在玻璃体腔内可见CD90和CD73阳性细胞。单次注射后1个月，玻璃体腔内可见存活的MSC。我们的结果证实了BLI是检测注射到眼球内的MSC的一种有用和可靠的方法。
Bone marrow-derived mesenchymal stromal cell (MSC) therapy is a promising treatment for several degenerative ocular diseases; however, no reproducible method of monitoring these cells into the eye has been established. The aim of this study was to describe successful bioluminescence imaging (BLI) to detect viable luciferase-expressing MSC in the eye. Human donor MSC in culture were transduced with 50 μl luciferase lentiviral vector (three viral particles/cell) prior to intraocular injection. Twenty-one right eyes of 21 rabbits were evaluated through BLI after receiving 1 × 106 luciferase-expressing MSC intravitreally. Contralateral eyes were injected with vehicle (phosphate-buffered saline (PBS)) and were used as controls. At seven different time points (1 h to 60 days), d-luciferin (40 mg/ml, 300 μl PBS) was injected in subsets of six enucleated eyes for evaluation of radiance decay through BLI analysis. CD90 and CD73 immunofluorescence was studied in selected eyes. Eyes injected with MSC showed high BLI radiance immediately after d-luciferin injection and progressive decay until 60 days. Mean BLI radiance measures from eyes with luciferase-expressing MSC were significantly higher than controls from 8 h to 30 days. At the thirtieth day, positive CD90- and CD73-expressing cells were observed only in the vitreous cavity of eyes injected with MSC. Viable MSC were identified in the vitreous cavity 1 month after a single injection. Our results confirmed BLI as a useful and reliable method to detect MSC injected into the eye globe.
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影响因子：
5.2
作者：
Manuguerra-Gagne, Renaud;Boulos, Patrick R.;Roy, Denis-Claude
通讯作者：
Roy, Denis-Claude
影响因子：
4.3
作者：
Miteva K;Van Linthout S;Volk HD;Tschöpe C
通讯作者：
Tschöpe C
通过系统施用的骨髓衍生的间充质干细胞对视网膜伤口愈合的调节。
影响因子：
--
作者：
Chung JK;Park TK;Ohn YH;Park SK;Hong DS
通讯作者：
Hong DS
影响因子：
1
作者：
Castanheira, Paula;Torquetti, Leonardo;Goes, Alfredo Miranda
通讯作者：
Goes, Alfredo Miranda
DOI：
10.1007/s00259-008-0751-z
发表时间：
2008-10-01
影响因子：
9.1
作者：
Hwang, Do Won;Jang, Sung June;Lee, Dong Soo
通讯作者：
Lee, Dong Soo