诱导脐血单个核细胞分化为树突状细胞的实验研究

Differentiation into dendritic cells induced from cord blood mononuclear cytes

摘要:

目的：诱导脐血源树突状细胞对恶性血液病脐血移植后的过继免疫治疗十分重要。诱导条件对树突状细胞产率影响很大。本研究观察粒细胞-巨噬细胞集落刺激因子(GM-CSF)、IL-4及TNF-α分阶段诱导脐血单个核细胞来源树突状细胞的效果。方法：通过Ficoll-Hypaque法分离脐血单个核细胞，以GM-CSF、IL-4及TNF-α分阶段诱导脐血来源树突状细胞的生成，培养12 d收获细胞，以光镜和电镜观察其细胞形态学特点，以流式细胞术检测其表面标记。结果：经培养12 d，每2×10 6脐血单个核细胞收获树突状细胞数为(0.46±0.13)×106，CD80+、CD86+细胞比例分别为89.98%±4.32%、86.86%±7.17%，而CD1a+细胞比例43.16%±5.80%，经光镜和电镜下观察细胞具有典型的树突状细胞形态学特征。结论：通过GM-CSF、IL-4及TNF-α的树突状细胞培养体系和分阶段诱导法可高效率诱导脐血树突状细胞。［中国当代儿科杂志，2004, 6(5): 377-380］

Abstract:

OBJECTIVE: The production of dendritic cells depends on the different culture conditions. It is necessary to establish a high effective dendritic cells culture system for high production. This study aims to explore the feasibility of programmable expansion of dendritic cells from cord blood mononuclear cytes by the induction of combination of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-4 and TNF-α. METHODS: Cord blood mononuclear cytes were separated by density centrifugation on Ficoll-Hypaque grad. Dendritic cells generated from cord blood mononuclear cytes were produced by the induction of a combination of GM-CSF, IL-4 and TNF-α. After 12 days of induction, the morphology of the harvested cells was observed under the light and electron microscopes, and the surface expression was examined using a flow cytometer. RESULTS: On the 12th day of induction, ( 0.46± 0.13)×10 6 dendritic cells were harvested from 2×10 6 cord blood mononuclear cytes. The proportion of CD80 +, CD86 + and CD1a + positive cells were 89.98%± 4.32%, 86.86%± 7.17% and 43.16%± 5.80% respectively. The expanded cells presented a typical dendritic cell morphology. CONCLUSIONS: The programmable culture system with combination of GM-CSF, IL-4 and TNF-α can effectively induce and expand cord blood dendritic cells.