Gasco Samanta, Rando Amaya, Zaragoza Pilar, García-Redondo Alberto, Calvo Ana C, Osta Rosario
Laboratory of Genetics and Biochemistry (LAGENBIO), Veterinary Faculty of Zaragoza, Aragon Institute of Food Agriculture, University of Zaragoza, Zaragoza, Spain
Biochemistry Department, Health Research Institute, Madrid, Spain
Comp Med. 2017 Feb 1;67(1):22-27. Epub 2017 Jan 12.
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.
确定实验所需合适且数量最少的对照小鼠是实验设计中的关键步骤。鉴于小鼠造血细胞群体存在各种与年龄相关的变化，这一决策对于长期研究造血系统尤为重要。在此，我们使用流式细胞术连续监测造血干细胞、普通淋巴样祖细胞和普通髓样祖细胞的频率，并采用逆转录聚合酶链反应（RT-PCR）分析来研究野生型B6SJL遗传背景的雄性和雌性小鼠中控制造血过程的Ly6a（Sca1）、Slamf1、Ikzf1和Cebpa这4个基因的水平。这些分析揭示了不同发育阶段的未成熟和成熟小鼠在细胞水平和mRNA水平上存在许多差异。总之，尽管有必要尽可能减少小鼠数量以减少动物使用并满足动物福利要求，但我们的研究结果所显示的众多差异凸显了在对造血系统细胞进行研究时，为所选的每个时间点都设立对照的必要性。在比较小鼠发育的未成熟和成熟阶段时，这种必要性尤为关键。
