中国医学科学院学报2012，Vol.34Issue(1)：19-24,6.DOI:10.3881/j.issn.1000-503X.2012.01.004
CD34+CD38- Lin -细胞群单个细胞培养的初步研究
A Preliminary Study on the Culture of Single Hematopoietic Stem Cell
摘要
Abstract
Objective To investigate the biological behavior including survival and proliferation of CD34 + CD38 - Lin - cells when they are cultured at single cell level. Methods Purified umbilical cord blood CD34 + CD38 - Lin - cells were separated at single cell level in 96-well plates using flow cytometry for four goups: control group ( CD34 + CD38 - Lin - cell plus stem cell medium) , Shh group ( CD34 + CD38 -Lin - cell plus stem cell medium and Shh) , BMP-4 group ( CD34 + CD38 - Lin - cell plus stem cell medium and BMP-4) , Jagged-1 group ( CD34 + CD38 - Lin - cell plus stem cell medium and Jagged-1). Methylcel-lulose medium was used in the colony-forming experiment which was also in four groups as previously. The number of cells and colony-forming units in each well for the four groups was evaluated at different time points ( day 1,3,7) with fluorescence microscopy counting method. Results Division of single cell was observed to be amplified in all of these groups from day 3. And meanwhile, after 1-week culture, the survival rates for the treated groups were all higher than the control group ( Jagged-1 group > BMP-4 group > Shh group > control) , while the cell number in each well was also highest in the Jagged-1 group (Jagged-1 group > BMP-4 group > control). The number of wells with a cell number of zero was significantly fewer in all treated groups ( especially the Jagged-1 group) than in the control group; meanwhile, the number of wells with a cell number higher than 17 was evidently higher in all the treated groups ( especially the BMP-4 group) more than controls. Colony-forming units for erythroid ( BFU-E ) , granulocyte ( CFU-G) , macrophage ( CFU-M) , and granulocyte macrophage ( CFU-GM ) were observed for all of these experimental groups, and there was no significant difference between the four experimental groups. Conclusions CD34 + CD38 - Lin - cell can achieve the survival, self-renewal and proliferation when cultured at single cell level, and the adding of Shh, BMP-4, and Jagged-1 can enhance such capabilities. However, CD34 + CD38 - Lin - cell can only maintain cell totipotency in its niche.关键词
CD34+CD38-Lin-细胞/Shh/BMP-4/Jagged-1/自我更新/增殖/微环境Key words
CD34 + CD38 - Lin - cell/ Shh/BMP-4/Jagged-1/self-renewal/ proliferation/microenvironment分类
生物科学引用本文复制引用
韩有金,齐军元,邱录贵..CD34+CD38- Lin -细胞群单个细胞培养的初步研究[J].中国医学科学院学报,2012,34(1):19-24,6.基金项目
天津市科技计划项目(09JCYBJCH11100)和教育部博士点新教师基金(200800231131) （09JCYBJCH11100）
