新分裂干细胞的鉴定

Q4 Biochemistry, Genetics and Molecular Biology
Dmitry I Maltsev, Oleg V Podgorny
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引用次数: 0

摘要

静止和增殖状态之间的交替能力是许多类型体细胞干细胞的显著特征。静止和增殖状态之间的平衡对干细胞在整个生命周期中的维持至关重要,如果这种平衡被打破,可能导致干细胞池过早枯竭,并在生命后期丧失组织再生或更新能力。如何调节这种平衡,对干细胞研究和衰老生物学至关重要。由于缺乏稳健测定干细胞退出可逆细胞周期停滞的速率的方法,直到最近,对静止与增殖状态之间平衡的评估仍面临挑战。在这里,我们提出了一种简单的方法,用于检测经过长期静止后进入分裂周期的干细胞。该方法结合了胸腺嘧啶类似物5-溴-2'-脱氧尿苷(BrdU)和5-乙炔基-2'-脱氧尿苷(EdU)的累积标记和脉冲标记。在所讨论的标记方案中,只加入第二个标记--EdU--的细胞是新分裂细胞。建议的双重标记方法可对干细胞退出静止状态的速度进行定量评估,并可追溯新分裂干细胞的命运。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Identification of De Novo Dividing Stem Cells.

The ability to alternate between quiescent and proliferating states is a remarkable feature of many types of somatic stem cells. The balance between quiescent and proliferating states is vital for maintenance of stem cells over the lifespan, and its disturbance may lead to premature depletion of the stem cell pool and loss of the tissue regenerative or renewal capacity at later stages of life. The question on how this balance is regulated is of critical importance in stem cell research and biology of aging. Assessment of the balance between quiescent and proliferating states has remained challenged until recently due to the lack of approaches for robust determination of the rate at which stem cells exit reversible cell cycle arrest. Here, we propose a simple method for detection of those stem cells that have entered the division cycle after a prolonged period of quiescence.The method combines cumulative and pulse labeling with thymidine analogues 5-bromo-2'-deoxyuridine (BrdU) and 5-ethynyl-2'-deoxyuridine (EdU). In the discussed labeling scheme, cells that have incorporated only the second label, EdU, are de novo dividing cells. The suggested double labeling method provides quantitative assessment of the rate at which stem cells exit the quiescent state and allows the fates of de novo dividing stem cells to be traced.

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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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