主动脉-性腺-中肾区、卵黄囊和循环血来源的CD41^＋细胞分化潜能比较  

Differentiation potential of CD41^＋ cells derived from the mouse aorta- gonad- mesonephros region,yolk sac and embryonic circulating blood

作　　者：李思霆[1] 周军年[1] 陈海旭[1] 谢一帆[1] 贺文艳[2] 南雪[1] 岳文[1] 刘兵[2] 裴雪涛[1] 

机构地区：[1]军事医学科学院野战输血研究所全军干细胞与再生医学重点实验室,北京100850 [2]军事医学科学院附属医院肿瘤学研究室

出　　处：《中华血液学杂志》2013年第10期887-892,共6页Chinese Journal of Hematology

基　　金：国家重点基础研究发展规划（重大科学研究计划）项目（2011CB964804）;国家自然科学基金（81101812）

摘　　要：目的比较小鼠胚胎不同造血组织来源的CD41^＋细胞向造血、间质、内皮细胞分化的潜能。方法取小鼠胚胎孕11d主动脉-性腺-中肾（AGM）区、卵黄囊（YS）和循环血（CB）来源的造血细胞经流式细胞术分选获得CD41^＋细胞，并检测CD41^＋细胞中CD45和c．kit的表达；采用IL-3、骨形态发生蛋白4（BMP-4）预处理半固体培养法评估CD41^＋细胞造血分化潜能的差异；采用荧光免疫法检测CD41^＋细胞α-平滑肌肌动蛋白（α-SMA，间质细胞标志）的表达；采用向内皮细胞分化诱导的培养条件观察其内皮分化潜能。结果在AGM区、YS、CB来源的CD4l^＋细胞中，CD45^＋细胞比例分别为51．9％、45．8％、22．2％，c-kit^＋细胞比例分别为40．0％、39．6％、36．2％。在IL-3刺激后，与未刺激组相比AGM、YS、CB来源的CD41’细胞总的造血细胞集落形成数量均显著增加[（14．1=k1．9）对（1．2＋0．2）；（32．4士1．1）对（18．4士2．2）；（41．84-0．9）对（10．44-1．8）]（P〈0．01）。BMP-4刺激后，与未刺激组相比：AGM、YS来源的CD41’细胞生成CFU-Mix的数量下降[（0．54-0．6）对（3．2士0．8）；（1．3＋0．7）对（7．4土1．7）]（P〈0．01）；但CB来源的CD41^＋细胞生成CFU．Mix的数量无明显变化[（2．54-0．5）对（3．94-1．5）]（P〉0．01）。AGM、YS来源的CD41^＋细胞高表达α-SMA，但CB来源的CD41^＋细胞低表达d-SMA。结论小鼠胚胎不同造血组织来源的CD41^＋细胞在免疫表型、造血细胞集落形成及细胞因子刺激应答方面存在明显差异。Objective To compare the differentiation ability difference of hematopoietic, mesenchymal and endothelial potential between CD41^＋cells derived from the mouse aorta-gonad-mesonephros （AGM） region, yolk sac （YS） and embryonic circulating blood （CB）. Methods CD41^＋cells were sorted from AGM, YS and CB. The CD45 and c-kit expression were studied in CD41^＋cells by flow cytometry. IL-3 and bone morphogenetic protein 4 （BMP-4） treatment together with semi solid culture were used to assess hematopoietic potential difference of CD41^＋ cells. Immunofluorescence staining of α-SMA was used to assess mesenchymal potential difference. The endothelial cell induction system was used to assess endothelial potential difference. Results The proportions of CD45 ^＋ cells in CD41^＋population were 51.9% （AGM）, 45.8% （YS） and 22.2% （CB）, respectively, while those of e-kit^＋ cells were 40.0% （AGM）, 39.6% （YS） and 36.2% （CB）, respectively. After stimulated by IL-3 factor, the number of total colonies increased in all three groups-derived CD41^＋ cells compared to that of unstimulated group [（14.1士1.9） vs （1.2士0.2）, （32.4士1.1） vs （18.4士2.2） and （41.8士0.9） vs （10.4士1.8）], （P〈0.01）. After stimulated by BMP-4 factor, compared to unstimulated group, CFU-Mix colony number in CD41^＋ cells from AGM region and YS were significantly decreased [ （0.5士0.6） vs （3.2士0.8）, （1.3士0.7） vs （7.4士 1.7） ] （P〈0.01） , but there was no difference in CB group [ （2.5士0.5） vs （3.9士 1.5） ] （P〉0.01）. The mesenehymal marker ct-SMA was highly expressed in CD41 ^＋ cells from AGM region and YS, but lowlyexpressed in CD41^＋ cells from CB. Conclusions There are some differences between CD41 ^＋ cells in AGM region, YS and CB on hematopoietic cell surface marker expression, hematopoietic colony formation with IL-3 and BMP-4 stimulation.

关 键 词：造血 胚胎发育 白细胞介素3 骨形态发生蛋白质4 抗原 CD41 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]