| Analysing Temporal Dynamics of T Cell Division in vivo Using Ki67 and BrdU Co-labelling by Flow Cytometry
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Author:
Date:
2017-12-20
[Abstract] This protocol was developed to increase the richness of information available from in vivo T cell proliferation studies. DNA labelling techniques such as BrdU incorporation allow precise control of label administration and withdrawal, so that the division history of a population can be tracked in detail over long timeframes (days-weeks). Ki67 is expressed in the nucleus of dividing cells, and is retained for a short time (3-4 days) after division (Gossel et al., 2017); therefore acting as a molecular clock to identify cells that have recently divided. Combining these two ...
[摘要] 该协议的开发是为了增加从体内T细胞增殖研究中获得的信息的丰富性。 诸如BrdU掺入的DNA标记技术允许精确控制标记施用和撤回，从而可以在很长时间段（日 - 周）内详细地追踪群体的分裂历史。 Ki67在分裂细胞的细胞核中表达，并在分裂后短时间（3-4天）保留（Gossel等人，2017）。 因此充当分子时钟来识别最近分裂的细胞。 将这两种技术结合起来就可以整合来自个体细胞的当前和历史扩散信息。 这个数据随后可以用来通过拟合增殖的数学模型来探测种群动态（Gossel et al。，2017）。
【背景】量化T细胞死亡和分裂的动力学是一个重大的实验和计算挑战，但是这个信息对于我们理解一个健康的免疫系统如何发展和维持至关重要。 T细胞群体动态的体内分析有多种技术，但数据的解释是复杂的，结果可能是模型依赖的（Asquith et al。2002 ; De Boer和Perelson，2013）。许多这些分析基于流式细胞仪读数，通过利用多参数分析的潜力提供获得新颖见解的机会。 BrdU吸收与Ki67表达一起的分析结合是特别有用的，因为每个代表细胞分裂的独特参数。 BrdU是掺入分裂细胞DNA中的胸苷类似物，因此永久标记在标记期间已分裂的细胞（Tough和Sprent，1994; ...
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| In vivo BrdU Incorporation Assay for Murine Hematopioetic Stem Cells
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Author:
Date:
2013-11-05
[Abstract] Bromodeoxyuridine (BrdU) is a thymidine analog that is incorporated into DNA during the S-phase of the cell cycle. As such, BrdU incorporation can be used to quantify the number of cells that are in S-phase in the time period during which BrdU is available. The following protocol describes an in vivo BrdU incorporation assay as a measure of cell proliferation in adult murine hematopioetic stem cells (HSCs). Specifically, BrdU incorporation was analyzed for long-term HSCs (LT-HSCs, Lin-Sca-1+c-Kit+CD34-CD135-), Short-term HSCs (ST-HSCs, ...
[摘要] 溴脱氧尿苷（BrdU）是在细胞周期的S期期间掺入DNA的胸苷类似物。 因此，BrdU掺入可用于定量在BrdU可用的时间段内处于S期的细胞的数量。 以下方案描述了体内BrdU掺入测定作为成体鼠造血干细胞（HSC）中细胞增殖的量度。 具体地，分析BrdU掺入的长期HSC（LT-HSC，Lin-Sca-1 + c-Kit + CD34 - CD135 - ），短期HSC（ST-HSC，Lin-Sca-1 + c-Kit + CD34 和多能祖细胞（MPP，Lin-Sca-1 + c-Kit + CD34 + > CD135 + ）群体
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| In vivo BrdU Incorporation and Detection in Mouse
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Author:
Date:
2011-06-05
[Abstract] BrdU (Bromodeoxyuridine or 5-bromo-2’-deoxyuridine) is a synthetic nucleoside that is incorporated into DNA by proliferating cells. This protocol is to be used to incorporate and detect BrdU in murine plasma cells. The plasma cells described in this protocol are formed spontaneously in autoimmune mice (NZB/W mice). Modifications are most likely needed if users intend to label plasma cells in immunized mice.
[摘要] BrdU (溴脱氧尿苷 or 5-溴-2’-脱氧尿苷) 是一种人工合成的核苷酸，能通过增殖细胞插入DNA中. 这个方法是用来在鼠浆细胞中标记和检测BrdU . 在这个方法中的浆细胞是在自身免疫的小鼠(NZB/W mice)中自发产生的.如果想要在免疫过的小鼠中标记 浆细胞，修饰时最可能用到的.
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