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聚酰胺-胺树枝状聚合物偶联量子点用于原代培养间充质干细胞的高效标记。

Polyamidoamine dendrimer-conjugated quantum dots for efficient labeling of primary cultured mesenchymal stem cells.

机构信息

Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Yoshida-shimoadachi, Sakyo-ku, Kyoto 606-8501, Japan.

出版信息

Biomaterials. 2011 Oct;32(28):6676-82. doi: 10.1016/j.biomaterials.2011.05.076. Epub 2011 Jun 22.

Abstract

Monitoring of cells in vivo after transplantation could supply important information for determining the efficacy of stem cell therapy. The use of quantum dots (QDs) has several advantages for in vivo imaging, such as remarkable resistance to photo bleaching, high fluorescence efficiency, and size-tunable emission. After they are taken up by cells via endocytosis, QDs lose their fluorescence intensity in endosomes/lysosomes at low pH because the intensity cannot survive under acidic conditions. Moreover, the amount of QD uptake by mesenchymal stem cells (MSCs) is extremely small. Therefore, for effective labeling of MSCs and long observation of MSCs labeled by QDs in vivo, it is essential both to increase cellular uptake of QDs and to promote endosomal escape into the cytosol. The polyamidoamine (PAMAM) dendrimer had plenty of cationic charge, which promoted cellular uptake though electrostatic interactions, and a "buffering capacity," which enhanced endosomal escape into the cytosol. In this study, QDs were modified with PAMAM dendrimer for the efficient labeling of MSCs by QDs. The uptake efficiency and cytosolic distribution of QDs in primary cultured MSCs were increased by the modification of the PAMAM dendrimer. The fluorescence intensity in MSCs labeled by PAMAM dendrimer-conjugated QDs lasted for a longer time in harvested culture plates or in cell-transplanted mice than that in MSCs labeled by non-conjugated QDs.

摘要

移植后细胞的体内监测可为确定干细胞治疗的疗效提供重要信息。量子点(QD)的使用在体内成像中有几个优点,例如对光漂白有显著的抗性、高荧光效率和可调的发射尺寸。当 QD 通过内吞作用被细胞摄取后,由于强度不能在酸性条件下存活,它们在低 pH 值的内体/溶酶体中失去荧光强度。此外,间充质干细胞(MSC)摄取 QD 的量非常少。因此,为了有效地标记 MSC 并在体内长时间观察被 QD 标记的 MSC,增加 QD 的细胞摄取并促进内体逃逸到细胞质是至关重要的。聚酰胺-胺(PAMAM)树枝状大分子具有大量的正电荷,通过静电相互作用促进细胞摄取,并具有“缓冲能力”,增强内体逃逸到细胞质。在这项研究中,PAMAM 树枝状大分子修饰了 QD,以实现 QD 对 MSC 的有效标记。通过 PAMAM 树枝状大分子的修饰,提高了原代培养 MSC 对 QD 的摄取效率和细胞质分布。与非共轭 QD 标记的 MSC 相比,PAMAM 树枝状大分子共轭 QD 标记的 MSC 的荧光强度在收获的培养板或细胞移植的小鼠中持续时间更长。

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