组织培养用于小鼠胚胎AGM区造血发育的研究  

Analyzing Hematopoietic Development of Mouse Embryonic AGM Region by Tissue Culture

作　　者：陈冬波[1] 高娇[2] 李专[1] 贺文艳[1] 刘兵[1] 

机构地区：[1]军事医学科学院附属医院肿瘤学研究室,北京100071 [2]解放军第306医院检验科,北京100101

出　　处：《中国实验血液学杂志》2011年第5期1195-1199,共5页Journal of Experimental Hematology

摘　　要：本研究通过建立组织培养方法探讨小鼠胚胎主动脉-性腺-中肾区(AGM)区造血发育规律。选取小鼠胚胎AGM区作为实验对象,体外组织培养2天后,应用集落形成实验观察髓系祖细胞的数量,应用体内移植考察CFU-S11和造血干细胞的发育。结果表明:组织培养后的AGM区细胞仍然具有形成髓系集落的能力;每个胚胎期10.5天(E10.5)AGM区在致死剂量照射的成年小鼠脾脏中可形成10.8±3.5个CFU-S11。更重要的是,经组织培养的E10.5-E11.5 AGM区能高比例(85.7%)、高嵌合〔(51.12±21.17)%〕、长期(>4个月)重建受致死剂量照射的成年小鼠的造血系统,在受鼠的外周血、骨髓、脾脏、胸腺中均能检测到供体来源的淋系或髓系祖细胞嵌合。结论:应用本研究建立的组织培养体系可对正常或基因突变胚胎AGM区中的多种造血起始细胞(尤其是造血干/祖细胞)进行发育动力学研究。To analyze hematopoietic kinetics of mouse embryonic aorta-gonad-mesonephros（AGM） region,an in vitro tissue culture method was developed in this study,partly based on previous reports.After 2 days of tissue culture,a significant number of erythromyeloid progenitors,as quantitated by colony forming assay was detected in the AGM region.Moreover,the cells from cultured E10.5 AGM region could generate 10.8±3.5 colony-forming unit in spleen（CFU-S） per tissue on average.Transplantation of cultured E10.5-E11.0 AGM cells resulted in efficient（85.7% repopulated） and long-term（4 months） reconstitution of lethally irradiated adult recipients with remarkable chimerism（（51.12±21.17）%）.The multilineage contribution of donor cells was validated by significant engraftment of myeloid and/or lymphoid cells in peripheral blood,bone marrow,spleen and thymus of recipients.Taken together,the tissue culture method can enable us to manipulate the AGM region in vitro,fulfilling a systematic evaluation of developmental kinetics of various hematopoietic precursor cells,particularly HSC,in normal and mutant mid-gestation mouse embryos.

关 键 词：AGM区 造血干细胞 CFU-C CFU-S 组织培养 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]