Department of Emergency and Critical Care Medicine, Shanghai Changzheng Hospital, The Second Military Medical University, Shanghai 200003, China.
Fujian Provincial Key Laboratory on Hematology, Fujian Institute of Hematology, Fujian Medical University Union Hospital, Fuzhou, China.
Biosci Rep. 2018 Feb 21;38(1). doi: 10.1042/BSR20171589. Print 2018 Feb 28.
To evaluate the contributions of cellular memory mechanisms to hematopoietic stem/progenitor cell (HSPC) senescence. HSPCs (LinCD117, hereafter referred to as HSPC) were separated from young (6-week-old) and aged (18-month-old) mice using Magnetic Activated Cell Sorting (MACS). Cell cycle distribution of HSPCs was determined using flow cytometry. The mixed colony forming unit (CFU-Mix) assay was used to study the HSPCs' ability to proliferate. The mRNA expression levels of cellular memory-implicated PCG family (enhancer of zeste homolog 2 (Ezh2), B lymphoma mo-MLV insertion region 1 (Bmi-1), embryonic ectoderm development (Eed), melanoma nuclear protein 18 (Mel18), Mph1/polyhomeotic-like protein 1 (Rae-28)) and Trithorax group (TrxG) family (mixed lineage leukemia (Mll), thioredoxin (Trx)) were determined by quantitative real-time PCR. We obtained highly purified populations of mouse HSPCs (LinCD117) (92.2 ± 4.5% CD117). The percentage of HSPCs was significantly higher in older mice compared with younger control mice and the percentage of SA-β-galactosidase positive cells was significantly higher in HSPCs isolated from older mice (<0.05). The percentage of HSPCs in G/G was significantly higher in older mice compared with younger control mice (52.0 compared with 47.1%), indicating increased cell cycle arrest in senescent HSPCs. The amount of CFU-Mix was significantly decreased in aged group (13.8 compared with 40.0), indicating a diminished ability to proliferate in senescent HSPCs. gene mRNA expression was significantly lower in HSPCs from older mice compared to younger controls, while mRNA expression was significantly higher in HSPCs from older mice (<0.05). The expression of genes associated with cellular memory is altered in senescent (Lin CD117) HSPCs, which may affect the potential plasticity of aged hematopoietic stem cells (HSCs) and thereby contribute to senescence-associated disease processes.
为了评估细胞记忆机制对造血干细胞/祖细胞（HSPC）衰老的贡献。使用磁激活细胞分选（MACS）从年轻（6 周龄）和年老（18 月龄）小鼠中分离 HSPC（LinCD117，以下简称 HSPC）。使用流式细胞术确定 HSPC 的细胞周期分布。混合集落形成单位（CFU-Mix）测定法用于研究 HSPC 的增殖能力。通过定量实时 PCR 确定细胞记忆相关 PCG 家族（增强子的锌指蛋白 2（Ezh2）、B 淋巴瘤 mo-MLV 插入区 1（Bmi-1）、胚胎外胚层发育（Eed）、黑素瘤核蛋白 18（Mel18）、Mph1/同源盒样蛋白 1（Rae-28））和 Trithorax 组（TrxG）家族（混合谱系白血病（Mll）、硫氧还蛋白（Trx））的 mRNA 表达水平。我们获得了高度纯化的小鼠 HSPC（LinCD117）群体（92.2±4.5% CD117）。与年轻对照组相比，老年小鼠中 HSPC 的比例显著更高，并且从老年小鼠中分离的 HSPC 中 SA-β-半乳糖苷酶阳性细胞的比例显著更高（<0.05）。与年轻对照组相比，老年小鼠中 HSPC 的 G/G 期比例显著更高（52.0 与 47.1%），表明衰老 HSPC 中的细胞周期停滞增加。老年组 CFU-Mix 的数量显著减少（13.8 与 40.0），表明衰老 HSPC 中增殖能力下降。与年轻对照组相比，老年小鼠 HSPC 中的 基因 mRNA 表达显著降低，而 基因 mRNA 表达显著升高（<0.05）。衰老（Lin CD117）HSPC 中与细胞记忆相关的基因表达发生改变，这可能影响老年造血干细胞（HSC）的潜在可塑性，并从而有助于衰老相关疾病过程。
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