脐血造血干/祖细胞体外扩增及移植动物模型分析  

Analysis of expansion of hematopioetic stem/progenitor cells from cord blood in vitro and engraftment in animal model of transplantation

作　　者：胡嘉波[1] 许文荣[1] 张锡然[2] 朱伟[1] 孙晓春[1] 顾可梁[1] 许化溪[1] 

机构地区：[1]江苏大学医学技术学院,江苏镇江212001 [2]南京师范大学生命科学院,南京210097

出　　处：《临床检验杂志》2004年第1期6-8,共3页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省自然科学基金项目 (BK2 0 0 2 0 0 6);江苏省卫生厅重点基金项目 (H95 0 9);江苏省教育厅自然科学基金项目( 0 2KJB3 10 0 0 2 )

摘　　要：目的　探讨脐血造血干 /祖细胞体外扩增和体内重建造血的潜能。方法　利用造血干 /祖细胞培养、体外扩增、移植动物模型等技术 ,研究不同比密ficoll分离液分离的脐血造血干 /祖细胞的造血活性。结果　比密 1.0 6 4 g/ml分离液分离的 10 5个脐血MNC中 ,CFU GM集落数为 373± 2 89,BFU E为 12 1± 70。在G3(GM CSF和IL 3融合蛋白 )、IL 6、TPO(thrombopoietin)作用下 ,1.0 6 4 g/ml分离液分离的脐血造血干 /祖细胞在体外扩增 14d ,CFU GM扩增 5 2 .2倍。给经 8.5Gy致死剂量照射的小鼠输注 1.0 6 4 g/ml分离液分离的脐血造血干 /祖细胞 ,体内产生的脾结节 (CFU S)是输注 1.0 77g/ml分离液分离细胞的 2 .2倍。结论　比密 1.0 6 4 g/ml分离液分离的脐血造血干 /祖细胞 ,在体外具有较高的增殖。Objective To investigate the expansion of hematopoietic stem/progenitor cells from cord blood in vitro and the potential of hematopoietic reconstitution in vivo.Methods By using a series of techniques, i.e., stem/progenitor cells culture, expansion in vitro and transplantation in animal model, the hematopoietic activity of fractionated cells in discontinuous Ficoll gradient was evaluated.Results After separation of l.064 g/ml Ficoll gradient, the number of CFU-GM were 373±289/105 MNC and BFU-E were 121±70/105 MNC.Following treated with cytokines (G3,IL-6,TPO) up to 14 days, the number of CFU-GM expanded 52.2-fold in vitro. The number of CFU-S in Balb/c mice which were irradiated by a lethal dose of 8.5 Gy and transfused subsequently with the hematopoietic stem/progenitor cells separated by 1.064 g/ml Ficoll gradient was 2.2-fo1d as many as that of separated by l.077 g/ml Fico11 gradient.Conclusions The hematopoietic stem/progenitor cells after 1.064 g/ml Ficoll density separation acquired higher ability of expansion and sustaining hematopoiesis in vitro and the potential of hematopoietic reconstitution in vivo.

关 键 词：脐血 造血干细胞移植 造血祖细胞移植 细胞体外扩增 动物模型 造血活性 

分 类 号：R457[医药卫生—治疗学] R-33[医药卫生—临床医学]