摘要
目的探讨脐血库脐血分离、保存方法及造血干祖细胞的体外培养条件。方法采用建立在密度梯度离心基础上的HES分离法对脐血进行有核细胞分离、冻存及粒巨系造血祖细胞集落(CFU-GM)培养。结果离心管收集分离和四联袋采集分离的方法均可获得较高的单个核细胞量,但四联袋采集分离在回收率和浓缩程度上更为优越,且粒巨系造血祖细胞(CFU-GM)含量丰富,10%DMSO冻存液保存复苏后仍可获得较高活率和CFU-GM。结论我们建立的HES分离结合密度梯度离心分离脐血造血干祖细胞以及脐血长期冻存的方法为广泛、大规模地建立脐血库提供了基础。
[Objective] To investigate the conditions on processing and cryopreservation of umbilic cord blood （UCB） and expansion of hematopoietic precursors ex vivo for wide establishment of UCB bank. [Methods] We designed the procedure based on HES density gradient centrifugation and compared two strategies for UCB collection and. processing. [Results] The two strategies could get enriched nucleated cells. The average yield of total nucleated cells and the volume reducing in closed-bag system was higher than that in centrifuge tubes, even the latter was more convenient and less expensive. The potent proliferative ability of hematopoietic stem and progenitor cells was detected by the clone forming unit granulocyte-macrophage （CFU-GM） culture of post-processing and thawing nucleated cells. The effect of cryopreservation was also well proved by the recovery of thawing cells. [Conclusion] The procedure based on HES density gradient centrifugation and the cryopreservation method we designed provided the foundation of an effective and convenient procedure of collection, processing and preservation of UCB for UCB bank word-widely.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2007年第3期261-263,268,共4页
China Journal of Modern Medicine
关键词
脐血
造血干祖细胞
分离
保存
培养
umbilic cord blood bank
hematopoietic precursor
processing
preservation
