Age-Associated Differences in Hematopoietic Stem and Progenitor Cells of Mice.

Comparative medicine Pub Date : 2017-02-01 Epub Date: 2017-01-12
Samanta Gasco, Amaya Rando, Pilar Zaragoza, Alberto García-Redondo, Ana C Calvo, Rosario Osta
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Abstract

Establishing the appropriate yet minimal number of control mice for experiments is a critical step in experimental design. This decision is particularly important regarding the study of the hematopoietic system over time, given various age-associated changes in murine hematopoietic cell populations. Here we used flow cytometry to serially monitor the frequencies of hematopoietic stem cells, common lymphoid progenitor cells, and common myeloid progenitor cells and RT-PCR assays to study the levels of Ly6a (Sca1), Slamf1, Ikzf1, and Cebpa—4 genes that control the hematopoietic process—in wildtype male and female mice with a B6SJL genetic background. These analyses revealed many differences, both at the cellular and mRNA levels, between immature and mature mice at various developmental stages. In conclusion, although it is necessary to minimize the number of mice possible insofar as possible to reduce animal use and meet animal welfare requirements, the numerous differences shown by our findings highlight the need to establish controls for every time point selected for the study of the hematopoietic system cells. This need is especially crucial when comparing immature and mature stages of mouse development.

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小鼠造血干细胞和祖细胞与年龄有关的差异
在实验设计中,确定适当但数量最少的对照小鼠是关键的一步。考虑到小鼠造血细胞群中与年龄相关的各种变化,这一决定对于随时间变化的造血系统研究尤为重要。在这里,我们使用流式细胞仪连续监测造血干细胞、普通淋巴祖细胞和普通髓系祖细胞的频率,并使用 RT-PCR 分析法研究控制造血过程的 Ly6a(Sca1)、Slamf1、Ikzf1 和 Cebpa-4 基因的水平--野生型雄性小鼠和遗传背景为 B6SJL 的雌性小鼠。这些分析揭示了未成熟小鼠和成熟小鼠在不同发育阶段的细胞和 mRNA 水平上的许多差异。总之,尽管有必要尽可能减少小鼠的数量,以减少动物的使用并满足动物福利的要求,但我们的研究结果所显示的众多差异突出表明,有必要为研究造血系统细胞所选择的每个时间点建立对照。在比较小鼠发育的未成熟阶段和成熟阶段时,这种必要性尤为重要。
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