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目的探讨培养树突状细胞(DC)的最佳方法和合理的培养时间。方法取大鼠的骨髓细胞分别加入4种细胞因子重组大鼠粒细胞巨噬细胞集落刺激因子(rrGMCSF)5μg/L,重组大鼠白细胞介素4(rrIL4)5μg/L,重组大鼠肿瘤坏死因子α(rrTNFα)10μg/L,重组大鼠γ干扰素(rrγIFN)20μg/L培养2周,并用PE标记的抗大鼠CD86单克隆抗体检测它的成熟度。结果从第7天开始细胞周围开始逐渐伸出突起,第13天以后突起5～6支左右,且长度逐渐缩短,成熟的树突状细胞伸出长短不等的突起,类似神经细胞的树突。60只大鼠培养所得的树突状细胞平均为(1.18±0.21)×107第7、11、13、15天的CD86单抗的阳性率分别为30%、80%、92%、94%,随着时间的延长,它的阳性率不断增加,尤以第1周到第2周之间增长的特别明显,但2周以后,树突状细胞的成熟度无明显提高。结论应用骨髓法来培养树突状细胞加入细胞因子rrGMCSF、rrIL-4、rrTNF-α、rr-γ-IFN培养2周,可得到成熟的树突状细胞。
Objective To explore the best method of culturing dendritic cells (DC) and the reasonable culture time. Methods The myeloid cells of rats were treated with 5 μg / L recombinant human granulocyte macrophage colony stimulating factor (rgGSCSF), 5 μg / L rrIL4 recombinant rat tumor necrosis (RrTNFα) 10μg / L, recombinant rat interferon γ (rrγIFN) 20μg / L for 2 weeks, and its maturation was detected by PE labeled anti-rat CD86 monoclonal antibody. Results From the 7th day began to gradually protrude around the cell protrusions, after the first 13 days protruding about 5 to 6, and the length gradually reduced, mature dendritic cells protruding protrusions of varying lengths, similar to the dendrites of nerve cells . The average number of dendritic cells cultured in 60 rats was (1.18 ± 0.21) × 107. The positive rates of CD86 monoclonal antibodies were 30%, 80%, 92% and 94% on days 7, 11, With the extension of time, its positive rate continues to increase, especially in the first week to the second week of growth was particularly evident, but after 2 weeks, dendritic cells did not significantly improve the maturity. Conclusion Mature dendritic cells can be obtained by culturing dendritic cells by adding bone marrow cells into rrGMCSF, rrIL-4, rrTNF-α and rr-γ-IFN for 2 weeks.