应用骨髓法培养树突状细胞的研究  

Culture of dentritic cells by bone marrow method

作　　者：吴舰宇[1] 宋春芳[1] 许评[1] 宋纯[1] 石于波[1] 刘锐[1] 

机构地区：[1]哈尔滨医科大学附属第一医院普外一科,150001

出　　处：《中华实验外科杂志》2005年第7期796-797,共2页Chinese Journal of Experimental Surgery

摘　　要：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 most rational procedures for dendritic cell (DC) culture.Methods Rat bone marrow cells were isolated and cultured for 2 weeks with attendance of four cytokines,rrGM-GSF,rrIL-4,rrTNF-αand rrγ-IFN,respectively.PE labeled anti-rat CD86 monoclonal antibody was introduced to detect the maturity of the cells.[WT5”HZ]Results Processes of the cells came into being at the 7th day and after the 13th day,the number of processes of one cell could reach up to 6 but with decreasing length.Fully matured DCs presented processes with various length,which were quite similar to those of nerve cells.The number of cells obtained from the rat bone marrow was averaged to be (1.18± 0.21)×107.The positive rate for CD86 antibodies was 30%,80%,92% and 94% respectively corresponding to the 7th,11th,13th and 15th day,which indicated an increasing positive rate with the period of time.Conclusion Matured DCs could be obtained by isolation and 2-week culture of rat bone marrow with attendance of four cytokines,rrGM-GSF,rrIL-4,rrTNF-α and rrγ-IFN.

关 键 词：树突状细胞 巨噬细胞集落刺激因子 大鼠肿瘤坏死因子-Α 单克隆抗体检测 rTNF-α CD86 白细胞介素 Γ-IFN 培养时间 鼠粒细胞 因子重组 骨髓细胞 γ干扰素 神经细胞 细胞因子 5μg 成熟度 阳性率 种细胞 突起 

分 类 号：R392[医药卫生—免疫学]