Optimization of 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester for labeling human intervertebral disc cells in vitro
Optimization of 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester for labeling human intervertebral disc cells in vitro
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DOI：
10.3109/10520290009066489
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发表时间：
2000-05-01
影响因子：
1.6
通讯作者：
Hanley, EN
中科院分区：
文献类型：
--
作者：
Gruber, HE;Leslie, KP;Hanley, EN
我们已经评估了细胞内5-(和-6)-羧基荧光素二乙酸酯琥珀酰亚胺酯(CFSE)作为体外培养的人椎间盘细胞的跟踪标记的实用性。尽管5µM可用于标记细胞的全细胞荧光显微镜鉴定，但20µM更适合于石蜡包埋标记细胞的免疫细胞化学定位。在标记细胞的超微结构水平上可见致密的电子泡。在荧光显微镜下，也可以在整个细胞上观察到离散的囊泡标记。整个细胞保持良好的标记时间为6周。CFSE标记相对简单，对细胞无毒，无放射性。当组织工程研究需要确认免疫细胞化学阳性时，建议初步优化与特定细胞类型密切相关的细胞。
We have assessed the utility of an intracellular fluorochrome, 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), as a tracking label for human intervertebral disc cells in vitro. Although 5 mu M provides adequate intracellular labeling for whole cell fluorescent microscopic identification of labeled cells, 20 mu M was preferable for immunocytochemical localization of paraffin embedded labeled cells. Electron dense vesicles are seen at the ultrastructural level in labeled cells. Discrete vesicular labeling can also be observed in whole cell mounts viewed with fluorescence microscopy. Whole cells retain good label for 6 weeks. CFSE labeling is relatively easy, nontoxic to cells and nonradiocactive. Initial optimization of close with specific cells types is recommended when confirmation of positive immunocytochemistry is needed for tissue engineering studies.