Percoll密度离心富集慢性粒细胞白血病DC前体细胞

Enrichment of dendritic cells precursor from chronic myelogeous leukemia by Percoll density gradients

目的:建立一种有效、费用低的富集慢性粒细胞白血病树突状细胞(DC)前体细胞的方法。方法:Ficoll-Hypaque液常规分离慢性粒细胞白血病(CML)外周血单个核细胞,55%Percoll密度离心,比较Percoll分离后的低密度层细胞(B组)、高密度层细胞(C组)及未经Percoll分离细胞(A组)的细胞分类、诱导后CD1a、CD86表达。结果:55%Percoll混悬液分离后的B组细胞中早幼粒和中幼粒细胞所占百分比高于A组(P<0.05)和C组细胞(P<0.01),而淋巴细胞、嗜酸性粒细胞和嗜碱性粒细胞的百分比明显少于C组,较A组也有所减少。B组细胞经诱导14 d后CD1a、CD86阳性细胞百分率明显高于C组细胞和A组细胞(P<0.01)。结论:经55%Percoll混悬液分离外周血单个核细胞可提高DC诱导的比率,且该方法无毒、费用低、易操作、分离细胞数量大,可充分满足临床DC的回输需要。

Objective To set up an effective and low-price way for enriching dendritic cells precursor from chronic myelogeous leukemia by Percoll density gradients. Methods Peripheral blood mononuclear cells （PBMCs） were collected by separation through Ficoll-Hypaque, then PBMCs were separated by 55% Percoll gradients. The cell type and DCs expressing CDla, CD86 were detected in the high-density group （C） , low-density group （B） and non-Percoll separation group （A）. Results After separation of 55% Percoll, the percentage of promyelocytes and myelocytes in group B was obviously higher than those in group A and group C （P〈0.01）. Moreover the percentages of lymphocytes, eosinophilic granulocytes and basophilic granulocytes in group B were lower than those in group C and group A. The percentages of CDla and CD86 cells detected by FACS on 14th day in group B were higher than those in group A and group C （P〈0.01）. Conclusion The induced rate of dendritic cells is improved after the separation of 55% Percoll. This method is no-toxic, cheap , simple and can handle a large number of cells. It＇ s useful for clinical application.