Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan.
Small. 2011 Dec 2;7(23):3363-70. doi: 10.1002/smll.201101233. Epub 2011 Oct 14.
Fluorescent nanodiamond (FND) has excellent biocompatibility and photostability, making it well suited for long-term labeling and tracking of cancer and stem cells. To prove the concept, the exocytosis of FND particles (size ≈100 nm) from three cell lines--HeLa cervical cancer cells, 3T3-L1 pre-adipocytes, and 489-2.1 multipotent stromal cells--is studied in detail. FND labeling is performed by incubating the cells in a serum-free medium containing 80 μg mL(-1) FND for 4 h. No significant alteration in growth or proliferation of the FND-labeled cells, including the multipotent stromal cells, is observed for up to 8 days. Flow cytometric analysis, in combination with parallel cell doubling-time measurements, indicates that there is little (≈15% or less) excretion of the endocytosed FND particles after 6 days of labeling for both HeLa and 489-2.1 cells, but exocytosis occurs more readily (up to 30%) for 3T3-L1 preadipocytes. A comparative experiment with FND and the widely used dye, carboxyfluorescein diacetate succinimidyl ester, demonstrates that the nanoparticle platform is a promising alternate probe for long-term cell labeling and tracking applications.
荧光纳米金刚石(FND)具有极好的生物相容性和光稳定性,非常适合用于癌症和干细胞的长期标记和追踪。为了验证这一概念,详细研究了三种细胞系——宫颈癌细胞 HeLa、3T3-L1 前脂肪细胞和 489-2.1 多能基质细胞——中 FND 颗粒(大小约 100nm)的胞吐作用。通过将细胞在含有 80μg mL(-1) FND 的无血清培养基中孵育 4 h 来进行 FND 标记。在长达 8 天的时间内,FND 标记的细胞(包括多能基质细胞)的生长或增殖没有明显变化。流式细胞术分析结合平行细胞倍增时间测量表明,对于 HeLa 和 489-2.1 细胞,在标记 6 天后,内吞的 FND 颗粒的排泄量很少(约 15%或更少),但 3T3-L1 前脂肪细胞更容易发生胞吐作用(高达 30%)。与 FND 和广泛使用的染料羧基荧光素二乙酸琥珀酰亚胺酯的对比实验表明,纳米颗粒平台是用于长期细胞标记和跟踪应用的有前途的替代探针。
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