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使用超灵敏加速器质谱法对 C 胸腺嘧啶核苷标记的间充质干细胞进行体内示踪。

In vivo tracking of C thymidine labeled mesenchymal stem cells using ultra-sensitive accelerator mass spectrometry.

机构信息

Department of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-Ro, Gwangjin-Gu, 05029, Republic of Korea.

Advanced Analysis Center, Korea Institute of Science and Technology, Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul, 02792, Republic of Korea.

出版信息

Sci Rep. 2021 Jan 14;11(1):1360. doi: 10.1038/s41598-020-80416-9.

Abstract

Despite the tremendous advancements made in cell tracking, in vivo imaging and volumetric analysis, it remains difficult to accurately quantify the number of infused cells following stem cell therapy, especially at the single cell level, mainly due to the sensitivity of cells. In this study, we demonstrate the utility of both liquid scintillator counter (LSC) and accelerator mass spectrometry (AMS) in investigating the distribution and quantification of radioisotope labeled adipocyte derived mesenchymal stem cells (AD-MSCs) at the single cell level after intravenous (IV) transplantation. We first show the incorporation of C-thymidine (5 nCi/ml, 24.2 ng/ml) into AD-MSCs without affecting key biological characteristics. These cells were then utilized to track and quantify the distribution of AD-MSCs delivered through the tail vein by AMS, revealing the number of AD-MSCs existing within different organs per mg and per organ at different time points. Notably, the results show that this highly sensitive approach can quantify one cell per mg which effectively means that AD-MSCs can be detected in various tissues at the single cell level. While the significance of these cells is yet to be elucidated, we show that it is possible to accurately depict the pattern of distribution and quantify AD-MSCs in living tissue. This approach can serve to incrementally build profiles of biodistribution for stem cells such as MSCs which is essential for both research and therapeutic purposes.

摘要

尽管在细胞示踪、活体成像和体绘制分析方面取得了巨大进展,但仍然难以准确量化干细胞治疗后输注细胞的数量,特别是在单细胞水平上,这主要是由于细胞的敏感性。在这项研究中,我们展示了液体闪烁计数器 (LSC) 和加速器质谱 (AMS) 在研究放射性同位素标记脂肪间充质干细胞 (AD-MSCs) 在静脉 (IV) 移植后单细胞水平分布和定量的应用。我们首先展示了 C-胸苷(5 nCi/ml,24.2 ng/ml)的掺入对 AD-MSCs 的关键生物学特性没有影响。然后,我们使用这些细胞通过 AMS 追踪和量化通过尾静脉递送的 AD-MSCs 的分布,揭示了不同时间点不同器官每毫克和每个器官中存在的 AD-MSCs 的数量。值得注意的是,结果表明,这种高灵敏度的方法可以定量检测每毫克一个细胞,这有效地意味着 AD-MSCs 可以在各种组织中以单细胞水平检测到。虽然这些细胞的意义尚未阐明,但我们表明可以准确描绘活组织中 AD-MSCs 的分布模式并对其进行定量。这种方法可以为 MSCs 等干细胞建立生物分布概况,这对于研究和治疗目的都是必不可少的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a45a/7809063/89aeb27d2952/41598_2020_80416_Fig1_HTML.jpg

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