CAJ | 학술논문

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l1d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CD41+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±1.9)对(1.2±0.2);(32.4±1.1)对(18.4±2.2);(41.8±0.9)对(10.4±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[(0.5±0.6)对(3.2±0.8);(1.3±0.7)对(7.4±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[(2.5±0.5)对(3.9±1.5)](P＞0.01).AGM、YS来源的C D41+细胞高表达α-SMA,但CB来源的CD41+细胞低表达αt-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.
목적 비교소서배태불동조혈조직래원적CD41+세포향조혈、간질、내피세포분화적잠능.방법 취소서배태잉l1d주동맥-성선-중신(AGM)구、란황낭(YS)화순배혈(CB)래원적조혈세포경류식세포술분선획득CD41+세포,병검측CD41+세포중CD45화c-kit적표체;채용IL-3、골형태발생단백4(BMP-4)예처리반고체배양법평고CD41+세포조혈분화잠능적차이;채용형광면역법검측CD41+세포α-평활기기동단백(α-SMA,간질세포표지)적표체;채용향내피세포분화유도적배양조건관찰기내피분화잠능.결과 재AGM구、YS、CB래원적CD41+세포중,CD45+세포비례분별위51.9％、45.8％、22.2％,c-kit+세포비례분별위40.0％、39.6％、36.2％.재IL-3자격후,여미자격조상비AGM、YS、CB래원적CD41+세포총적조혈세포집락형성수량균현저증가[(14.1±1.9)대(1.2±0.2);(32.4±1.1)대(18.4±2.2);(41.8±0.9)대(10.4±1.8)](P＜0.01).BMP-4자격후,여미자격조상비:AGM、YS래원적CD41+세포생성CFU-Mix적수량하강[(0.5±0.6)대(3.2±0.8);(1.3±0.7)대(7.4±1.7)](P＜0.01);단CB래원적CD41+세포생성CFU-Mix적수량무명현변화[(2.5±0.5)대(3.9±1.5)](P＞0.01).AGM、YS래원적C D41+세포고표체α-SMA,단CB래원적CD41+세포저표체αt-SMA.결론 소서배태불동조혈조직래원적CD41+세포재면역표형、조혈세포집락형성급세포인자자격응답방면존재명현차이.
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 c-kit+ ceils 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±1.9) vs (1.2±0.2),(32.4±1.1) vs (18.4±2.2) and (41.8±0.9) vs (10.4±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±0.6) vs (3.2±0.8),(1.3±0.7) vs (7.4±1.7)] (P＜0.01),but there was no difference in CB group [(2.5±0.5) vs (3.9±1.5)] (P＞0.01).The mesenchymal marker α-SMA was highly expressed in CD41 + cells from AGM region and YS,but lowly expressed 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.