ERT2-Cre诱导的Rosa26R-YFP报告基因系统标记小鼠造血干祖细胞的条件和效率检测 |
| |
引用本文: | 罗琦,尹洁,孟歆怿,杨冰,雷蕾,孙琰,李丹丹,王瑾,王玺. ERT2-Cre诱导的Rosa26R-YFP报告基因系统标记小鼠造血干祖细胞的条件和效率检测[J]. 国际生物医学工程杂志, 2014, 37(5): 266-270 |
| |
作者姓名: | 罗琦 尹洁 孟歆怿 杨冰 雷蕾 孙琰 李丹丹 王瑾 王玺 |
| |
作者单位: | 300070,天津医科大学基础医学院 |
| |
基金项目: | 国家自然科学基金资助项目 |
| |
摘 要: | 目的 探索ERT2-Cre诱导的黄色荧光蛋白(YFP)报告基因系统标记小鼠造血干祖细胞的条件和效率.方法 通过基因型分型的方法,从Rosa26R-YFP小鼠与ERT2-Cre小鼠杂交得到的后代中,筛选出YFP及ERT2-Cre双阳性转基因杂交小鼠;采用免疫磁珠法从双阳性转基因小鼠骨髓细胞中富集c-kit+造血干祖细胞群进行体外培养,并用不同浓度4-羟基它莫西芬(OHT)作用不同时间以诱导YFP表达;流式细胞术检测c-kit+造血干祖细胞中YFP的表达量.结果 Rosa26R-YFP小鼠与ERT2-Cre小鼠的杂交后代共9只,采用基因型分型方法筛选出双阳性转基因小鼠5只;体外培养的c-kit+细胞部分形态发生变化,提示存在细胞分化;流式细胞术检测到c-kit+细胞中YFP表达量可达13.7%.结论 YFP报告基因系统在OHT诱导的ERT2-Cre作用下,能有效标记小鼠造血干祖细胞.
|
关 键 词: | 造血系统 造血干祖细胞 c-kit+细胞 ERT2-Cre Rosa26R-YFP报告基因系统 |
Conditions and efficiency of Rosa26R-YFP reporter gene system labeled mouse hematopoietic stem and progenitor cells induced by ERT2-Cre |
| |
Affiliation: | Luo Qi,Yin Jie,Meng Xinyi,Yang Bing,Lei Lei,Sun Yan,Li Dandan(College of Basic Medical, Tianjin Medical University, Tianjin 300070, China) |
| |
Abstract: | Objective To explore the conditions and efficiency of YFP-labeled mouse hematopoietic stem and progenitor cells induced by ERT2-Cre.Methods ERT2-Cre and YFP double-positive transgenic hybrid mice from all offsprings of Rosa-YFP mice and ERT2-Cre mice were obtained by genotyping,and c-kit+ cells were enriched in vitro using magnetic activated cell sorting from bone marrow cells of double-positive transgenic hybrid mice.The expression of YFP was induced by different concentrations of 4-hydroxytamoxifen (OHT) with different periods,and the expressing efficiency of YFP in c-kit+ cells was detected by flow cytometry.Results Five out of nine Rosa26R-YFP and ERT2-Cre mice crossbred progenies were YFP and Cre transgene double positive.Some morphological changes of c-kit+ cells cultured in vitro suggested the presence of cell differentiation.The expressing efficiency of YFP in c-kit + cells was up to 13.7%.Conclusions YFP reporter gene system can effectively mark mouse hematopoietic stem and progenitor cells induced by ERT2-Cre with the exist of OHT in vitro. |
| |
Keywords: | Hematopoietic system Hematopoietic stem and progenitor cells c-kit+ cells ERT2-Cre Rosa26R-YFP reporter gene system |
本文献已被 维普 万方数据 等数据库收录! |
|