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利用多光子显微镜和光可转化聚合物胶囊对个体人类神经细胞进行长期追踪。

Long-term tracing of individual human neural cells using multiphoton microscopy and photoconvertible polymer capsules.

机构信息

Vladimir Zelman Center for Neurobiology and Brain Rehabilitation, Skolkovo Institute of Science and Technology, Moscow 121205, Russia.

Federal Center for Brain and Neurotechnologies, Federal Medical-Biological Agency, Moscow 117997, Russia.

出版信息

J R Soc Interface. 2024 Oct;21(219):20240497. doi: 10.1098/rsif.2024.0497. Epub 2024 Oct 30.


DOI:10.1098/rsif.2024.0497
PMID:39471872
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11521627/
Abstract

The study of human neural cells, their behaviour and migration are important areas of research in the biomedical field, particularly for potential therapeutic applications. The safety of using neural cells in therapy is still a concern due to a lack of information on long-term changes that may occur. While current methods of cell tracing explore gene manipulations, we elaborate approaches to cell marking with no genetic interference. In this study, we present a novel method for labelling and tracking neural cells using cell-impregnatable photoconvertible polyelectrolyte microcapsules. These capsules demonstrated low cytotoxicity with no effect on the differentiation ability of the neural cells, maintained a high level of fluorescent signal and ability for tracing individual neural cells for over 7 days. The capsules modified with rhodamine- and fluorescein-based dyes were demonstrated to undergo photoconversion by both one- and two-photon lasers while being internalized by neural cells. The finding gives the possibility to select individual capsules inside multicellular structures like spheroids and tissues and alternate their fluorescent appearance. Thus, we can track individual cell paths in complex systems. This new method offers a promising alternative for studying neural cells' long-term behaviour and migration in complex systems such as three-dimensional cellular populations.

摘要

研究人类神经细胞及其行为和迁移是生物医学领域的重要研究领域,特别是对于潜在的治疗应用。由于缺乏关于可能发生的长期变化的信息,使用神经细胞进行治疗的安全性仍然令人担忧。虽然目前的细胞示踪方法探索基因操作,但我们详细介绍了不进行基因干扰的细胞标记方法。在这项研究中,我们提出了一种使用可细胞浸渍的光可转化聚电解质微胶囊标记和跟踪神经细胞的新方法。这些胶囊表现出低细胞毒性,对神经细胞的分化能力没有影响,保持高水平的荧光信号,并能够在超过 7 天的时间内追踪单个神经细胞。用罗丹明和荧光素基染料修饰的胶囊被证明可以通过单光子和双光子激光器进行光转化,同时被神经细胞内化。这一发现使得有可能在类球体和组织等多细胞结构中选择单个胶囊,并改变它们的荧光外观。因此,我们可以在复杂系统中跟踪单个细胞的路径。这种新方法为研究复杂系统(如三维细胞群体)中神经细胞的长期行为和迁移提供了一种很有前途的替代方法。

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Long-term tracing of individual human neural cells using multiphoton microscopy and photoconvertible polymer capsules.

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本文引用的文献

[1]
Multiple dyes applications for fluorescent convertible polymer capsules as macrophages tracking labels.

Heliyon. 2024-5-5

[2]
Autofluorescence is a biomarker of neural stem cell activation state.

Cell Stem Cell. 2024-4-4

[3]
Single-cell tracking as a tool for studying EMT-phenotypes.

Exp Cell Res. 2024-4-1

[4]
Hydrogen peroxide is not generated intracellularly in human neural spheroids during ischemia-reperfusion.

Free Radic Biol Med. 2024-2-20

[5]
Blue-shift photoconversion of near-infrared fluorescent proteins for labeling and tracking in living cells and organisms.

Nat Commun. 2023-12-19

[6]
Labeling and Tracking of Individual Human Mesenchymal Stromal Cells Using Photoconvertible Fluorescent Microcapsules.

Int J Mol Sci. 2023-9-4

[7]
Fluorescent polymer markers photoconvertible with a 532 nm laser for individual cell labeling.

J Biophotonics. 2023-6

[8]
Macrophage and Tracking via Anchored Microcapsules.

ACS Appl Mater Interfaces. 2022-11-23

[9]
Tracking distinct microglia subpopulations with photoconvertible Dendra2 in vivo.

J Neuroinflammation. 2021-10-15

[10]
Single-cell tracking reveals super-spreading brain cancer cells with high persistence.

Biochem Biophys Rep. 2021-9-9

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