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使用胸苷类似物推断细胞周期参数的方法。

Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues.

作者信息

Martí-Clúa Joaquín

机构信息

Unidad de Citología e Histología, Departament de Biologia Cel·lular, de Fisiologia i d'Immunologia, Facultad de Biociencias, Institut de Neurociències, Universidad Autónoma de Barcelona, Bellaterra, 08193 Barcelona, Spain.

出版信息

Biology (Basel). 2023 Jun 20;12(6):885. doi: 10.3390/biology12060885.

Abstract

Tritiated thymidine autoradiography, 5-bromo-2'-deoxyuridine (BrdU) 5-chloro-2'-deoxyuridine (CldU), 5-iodo-2'-deoxyuridine (IdU), and 5-ethynyl-2'-deoxyiridine (EdU) labeling have been used for identifying the fraction of cells undergoing the S-phase of the cell cycle and to follow the fate of these cells during the embryonic, perinatal, and adult life in several species of vertebrate. In this current review, I will discuss the dosage and times of exposition to the aforementioned thymidine analogues to label most of the cells undergoing the S-phase of the cell cycle. I will also show how to infer, in an asynchronous cell population, the duration of the G, S, and G phases, as well as the growth fraction and the span of the whole cell cycle on the base of some labeling schemes involving a single administration, continuous nucleotide analogue delivery, and double labeling with two thymidine analogues. In this context, the choice of the optimal dose of BrdU, CldU, IdU, and EdU to label S-phase cells is a pivotal aspect to produce neither cytotoxic effects nor alter cell cycle progression. I hope that the information presented in this review can be of use as a reference for researchers involved in the genesis of tissues and organs.

摘要

氚标记胸腺嘧啶核苷放射自显影、5-溴-2'-脱氧尿苷(BrdU)、5-氯-2'-脱氧尿苷(CldU)、5-碘-2'-脱氧尿苷(IdU)和5-乙炔基-2'-脱氧尿苷(EdU)标记已被用于识别处于细胞周期S期的细胞比例,并追踪这些细胞在几种脊椎动物胚胎期、围产期和成年期的命运。在本综述中,我将讨论上述胸腺嘧啶类似物的剂量和暴露时间,以标记大多数处于细胞周期S期的细胞。我还将展示如何在异步细胞群体中,根据一些涉及单次给药、连续核苷酸类似物递送以及两种胸腺嘧啶类似物双重标记的标记方案,推断G1、S和G2期的持续时间,以及生长分数和整个细胞周期的跨度。在此背景下,选择最佳剂量的BrdU、CldU、IdU和EdU来标记S期细胞是一个关键问题,既要避免产生细胞毒性作用,又不能改变细胞周期进程。我希望本综述中提供的信息能为参与组织和器官发生研究的人员提供参考。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/517b/10295744/55d2ea09f9dc/biology-12-00885-g001.jpg

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