人脐血造血干细胞在转瓶中的悬浮培养  

Suspended Culture of Human Umbilical Cord Blood Stem Cells in Spinner Bottle

作　　者：夏泉鸣[1] 康自珍[1] 谭文松[1] 

机构地区：[1]华东理工大学生物工程学院,上海200237

出　　处：《华东理工大学学报（自然科学版）》2004年第1期47-50,共4页Journal of East China University of Science and Technology

基　　金：国家863项目(102-12-08-01)

摘　　要：考察经分离纯化后的人脐血CD34+富集细胞在转瓶中的扩增特性,细胞因子为SCF+IL-3+IL-6。实验发现,无论24孔板静态培养还是转瓶悬浮培养,孔板和转瓶内总细胞数分别扩增了(658.7±98.0)倍和(107.3±15.0)倍,说明CD34+具有很大的扩增潜能。对比两种培养体系下细胞集落扩增倍数发现,培养初期孔板内的集落形成能力要大于转瓶,但到培养后期,孔板内集落形成能力下降速度远大于转瓶。孔板中的CFU-GM(粒细胞-巨噬细胞集落形成单位)扩增倍数在第14天为(5.6±0.4)倍,第21天为(1.7±0.5)倍,转瓶中在第14天为(2.1±0.7)倍,第21天为(1.8±0.5)倍。对比两者的CD34+扩增情况,孔板在第14天CD34+含量达到最大值,而到第21天则大大降低,而转瓶内CD34+含量仍可保持在原第14天的水平,转瓶内的CD34+细胞的扩增也比孔板保持较长时间。The expansion characteristics of CD34^+ selected cells were examined by two culture system: well flask and spinner bottle in the medium containing cytokine cocktails of SCF+IL-3+IL-6. Two culture systems were compared: static system in well flask and suspended culture system in spinner bottle. It was found that the CD34^+ selected cells presented a high proliferation potential. Within 3 weeks of culture period, the fold expansion of total cells in well flask and spinner bottle were (658.7±98.0) and (107.3±15.0) respectively. Compared with the colony forming potential, the cells in well flask were greater, but in the later culture period the colony number declined more sharply. From the day 14 to day 21, the fold expansion of CFU-GM in well flask decreased from (5.6±0.4) to (1.7±0.5 ) while the fold expansion in spinner bottle decreased from (2.1±0.7) to (1.8±0.5). The proportion of CD34^+ reached its maximum at day 14 in well flask and then decreased sharply while it can be maintained to day 21 in spinner bottle. The result showed that the CD34^+ cells cultured in spinner bottle can be maintained for longer time than those in well flask.

关 键 词：CD34^+细胞 转瓶 体外扩增 细胞因子 静态培养 悬浮培养 人脐血 造血干细胞 

分 类 号：Q813[生物学—生物工程]