低温冻存对脐血树突状细胞生物特性的影响

INFLUENCE OF BIOLOGICAL PROPERTIES OF CRYOPRESERVED DENDRITIC CELLS DERIVED FROM CORD BLOOD

目的 :观察低温冻存对脐血来源的树突状细胞 (DCs)培养生物特性的影响 ,为DCs的应用提供冻存方法。方法 :设未冻存脐血细胞 (UFCB)经增殖培养后产生的DCs(UFDCs)为对照组 ,来源于经低温冻存的脐血 (CPCB)的DCs和经低温冻存的DCs(CPDCs)为实验组 ,比较实验组与对照组DCs生物特性 ,包括细胞增殖量 ,不染色和染色的细胞形态 ,台盼蓝拒染回收率 (TBR) ;细胞表面抗原 ;混合淋巴细胞培养剌激指数 (SI)、抗毒 杀灭效应处理率(ER)。结果 :CPCB与UFCB比 ,形成DCs的时间迟 ,细胞增殖量、树突状细胞量、CD1a、CD83、HLA DR表达、SI、ER均低。CPDCs与UFDCs比 ,形成贴壁树突状细胞迟 ,细胞增殖量、树突状细胞量、CD1a、CD83、HLA DR表达、SI、ER均低。CPCB的TBR为 95.8% ,CPDCs的TBR为 88.7%。结论 :脐血细胞和经增殖培养的脐血DCs ,经低温冻存后复温培养 ,DCs的生物特性有一定的损伤 ,但基本功能仍比较完整 ,回收率均 >85%。

Aim: To provide experimental basis for clinical use of cryopreserved den dritic cells (DCs) by investigating the biological properties of cryopreserved D Cs derived from cord blood. Methods: The biological discrepancie s between unfrozen DCs (UFDC) derived from unfrozen cord blood (UFCB) and DCs fr om cryopreserved cord blood (CPCB) or cryopreserved DCs (CPDCs) were explored by morphological observation and immunophenotype analysis. Moreover,the stimulati ng index (SI) in mixed lymphocyte culture and cytotoxic eliminating rate (ER) w ere measured by MTT. Results: The TBR of CPCB and CPDCs were 9 5.8% and 88.7% respectively. Compared to UFCB, CPCB differentiated toward DCs in a delayed and decreased mode, displayed lower expression of CD1a, CD83 and HLA -DR, and had reduced SI and ER. Similarly, the CPDCs became adherent DCs later and grew less in number than UFDCs. After culture, the expression of CD1a, CD83 and HLA-DR as well as SI and ER were lower in CPDCs than those in UFDCs. Conclusion: Though the biological properties of CPCB and CPDCs were injur ed after cryopreservation, they still had relatively complete basic function. Th eir recoveries rate were >85%.