人CD34^+和CD34^-细胞在NOD/SCID小鼠体内的造血重建  

Hematopoietic Repopulating Ability of Human CD34^+ Cells and CD34^- Cells in NOD/SCID Mice

作　　者：金慧丽[1] 蔡海波[1] 杨施[1] 谭文松[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《生物工程学报》2008年第9期1588-1594,共7页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.20776043);上海市生物医药科技攻关项目(No.074319109)资助~~

摘　　要：利用非肥胖糖尿病型重症联合免疫缺陷型(NOD/SCID)小鼠模型,比较了新鲜及培养后的CD34+和CD34-细胞在体内植入及重建造血能力。从新鲜脐血及培养后的单个核细胞(MNC)中分离出CD34+和CD34-细胞,经尾静脉输注入经亚致死剂量照射的NOD/SCID小鼠体内,6周后处死存活的小鼠,取其骨髓、脾脏和外周血细胞,分别进行细胞表型分析、造血集落形成单位和人特异性基因的检测。经检测,输注CD34+细胞和混合细胞的小鼠,其体内CD45+细胞及人源各系血细胞的含量相近,两者均远远高于输注CD34-细胞的小鼠。输注培养后CD34-细胞的小鼠饲养6周后全部死亡,输注培养后CD34+细胞的小鼠存活率约为66.7%,而输注培养后混合细胞的小鼠全部存活,且在两组存活的小鼠体内均能检测到CD45+细胞及人源各系血细胞。结果表明:无论是新鲜还是培养后的CD34+细胞均具有在NOD/SCID小鼠体内植入和重建造血能力,而CD34-细胞不具有该能力,但CD34-细胞与CD34+细胞同时输注有助于提高小鼠的存活率,说明其对CD34+细胞在小鼠体内发挥植入和造血重建能力有一定的辅助作用。The hematopoietic repopulating ability of fresh and cultured CD34^＋ cells and CD34^- ceils derived from cord blood were compared by nonobese diabetic/severe combined immunodeficiency （NOD/SCID） mouse model. Fresh CD34^＋ cells and CD34^- ceils were isolated from fresh cord blood. Cultured CD34^＋ cells and CD34^- cells were separated from cultured mononuclear cells （MNC）. We transplanted these cells into sublethally irradiated NOD/SCID mice via the tail vein and sacrificed surviving mice after 6 weeks. The peripheral blood, spleen and bone marrow from each mouse were harvested for flow cytometry, colony-forming cells and human Alu sequences analyses. The proportions of CD45^＋ cells and human multilineage hematopoietic cells in NOD/SCID mice received CD34^＋ ceils were close to that in the mice received both CD34^＋ cells and CD34^- ceils, while it was significantly higher than that in the mice received CD34^- cells. Six weeks after transplantation, all the mice injected with cultured CD34^- cells dead. The survival rate of mice injected with cultured CD34^＋ cells was 66.7%. All of the mice injected with both cultured CD34^- and CD34^＋ cells survived. Moreover, CD45^＋ cells could be detected in all surviving mice, and human CD34, CD3, CD19, CD33 and CD71 antigen also could be detected on these CD45^＋ cells. The results showed that both fresh and cultured CD34^＋ cells had the capability of engraftment and hematopoiesis reconstitution, but CD34^-cells hadn＇t the ability. However, CD34^- cells had assistant effect on the hematopoietic repopulating ability of CD34^＋ cells.

关 键 词：CD34^-细胞 CD34^+细胞 NOD/SCID小鼠 造血重建能力 

分 类 号：Q813[生物学—生物工程]