不同来源血清或血浆培养条件下的树突状细胞表型和功能的比较

Comparison of phenotype and functions of dendritic cells in culture medium supplemented with different serum or plasma

目的:比较在添加100 mL/L胎牛血清(FBS)、20mL/L人AB血清和10 mL/L供者自体血浆3种不同培养条件下树突状细胞(DC)的表型和功能,为临床应用DC找到安全且有效的培养方法。方法:将外周血单个核细胞(PBMCs)分别在添加100 mL/L FBS,20 mL/L人AB血清,10 mL/L供者自体血浆培养条件下诱导成DC,在培养的第8天,利用流式细胞术(FCM)检测3种培养条件下DC的得率、纯度、表型和抗原捕获能力;用MTT的方法检测DC刺激同种异体淋巴细胞增殖的能力。结果:在DC集落数量、DC得率、DC纯度、抗原捕获能力和刺激淋巴细胞增殖的能力上,100 mL/L FBS培养组优于20 mL/L AB血清和10 mL/L自体血浆组,3组之间DC得率差异比较具有统计学的意义(P<0.05),而其他比较都无统计学的意义(P>0.05)。结论:利用供者自体血浆培养DC是一种安全有效的培养方法,符合细胞治疗规范,可替代异种血清或同种异体血清,成为临床级别DC培养的选择。

AIM： To find a safe and effective method of culturing dendritic cells （DCs） for clinical application by comparing the phenotype and functions of DCs in different culture mediums which are supplemented with 100 ml_/L fetal bovine serum （ FBS）, 20 mL/L human AB serum and 10 mL/L autologous plasma. METHODS： Peripheral blood mononuclear cells （PBMCs） were induced to DCs in 100 mL/L FBS, 20 mL/L human AB serum, and 10 mL/L autol- ogous plasma, respectively. On day 8, the purity, pheno- type and antigen uptake capability of DCs were examined by flow cytometry. The mixed lymphocyte reaction （MLR） was detected by MTr assay. RESULTS： Focused on DC aggre- gates, DC yield, DC purity, antigen uptake capability and ability of stimulating lymphocyte proliferation, the experiments showed that those of 100 mL/L FBS culture group were bet- ter than those of 20 mL/L human AB serum and 10 mL/L autologous plasma culture groups. There was no significant difference among these three culture groups （P 〉 0.05） except DC yield （ P 〈 0.05 ）. CONCLUSION： Autologous plasma is the best choice for DC cultivation among these three culture ways. It is a safe and efficient way for culturing DCs for clinical purpose.