Ficoll分离和MACs分选对T细胞BCL11b表达水平的影响

Effect on the Expreesion Level of BCL11b Gene in T Cells by Ficoll Isolation and MACs Selection

目的了解Ficoll分离和MACs分选过程对T细胞中BCL11b基因表达水平的影响情况。方法利用Ficoll分离3例正常人外周血单个核细胞(PBMC),进一步经MACs负性分选CD3+T细胞。分别收集培养0、6和20h的PBMC和CD3+T细胞,利用实时定量PCR方法检测各组样本中BCL11b基因的表达水平。结果经Ficoll分离的PBMC和MACs分选后CD3+T细胞在3个时间点BCL11b的表达水平无显著性差异(p>0.05),根据CD3+T细胞%标化后,各组BCL11b表达水平也均无显著性差异。结论Ficoll分离和MACs负性分选过程不影响T细胞中BCL11b表达水平,提示其不会影响T细胞的活化。

Objective To analysis the effect on the expression of BCLllb gene in T cells during the Ficoll isolation and MACs selcetion process. Methods Peripheral blood mononuclear cells （PBMC） from 3 cases of normal individuals were isolated by Ficoll and farther selected for CD3 ＋ T cells by MACs negative selection. The PBMC and CD3 ＋ T cells were cultured with different time point （0, 6 and 20 hours）. The expression level of BCLllB gene was detected by real - time PCR. Results The expression levels of BCL11 b in PBMC after Ficell isolation and in CD3 ＋ T cells after MACs selection at 3 time point of culture were not significantly different （p 〉 0. 05）, as well as after standardization with percentage of CD3 ＋ T cells. Conclusions There are not significant changed on the expression of BCL11b gene in T cells during Ficoll isolation and MACs negative selection proccess, suggesting that might not affect on the activation of T cells.