小鼠肝脏树突状细胞前体分离培养及影响其成熟因素的初步探讨  

A PRELIMINARY STUDY OF ISOLATION AND CULTURE OF DENDRITIC CELL PRECURSORS FROM MOUSE LIVER AND INFLUENCE FACTOR OF DC MATURATION

作　　者：张宏[1] 钟翠平[1] 成令忠[1] 顾云娣[1] 王国强[1] 

机构地区：[1]上海医科大学组织胚胎学教研室

出　　处：《中国组织化学与细胞化学杂志》1998年第3期8-11,共4页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：本次首次建立了从小鼠肝脏分离、培养、扩增肝树突状细胞（ｈｅｐａｔｉｃｄｅｎｄｒｉｔｉｃｃｅｌ，ＨＤＣ）前体的方法。采用经门静脉插管胶原酶二步循环灌流法及Ｐｅｒｃｏｌ不连续密度梯度离心法分离非肝细胞，得到含ＨＤＣ前体组分，将其分三组培养：第一组加入重组小鼠粒细胞／巨噬细胞集落刺激因子（ＭｒＧＭＣＳＦ），待培养９天后，将细胞转移至鼠尾胶原上继续培养。第二组加入ＭｒＧＭＣＳＦ培养，但细胞不转移。第三组为对照组，仅加完全培养液。结果显示：第一、二组细胞于培养４天可见有许多细胞集落形成，第一组细胞转移至鼠尾胶原上培养１天后，细胞集落减少，而培养液中ＤＣ密度明显增加，光镜和扫描、透射电镜观察证明培养ＨＤＣ纯度达９５％，第二组则无上述细胞释放现象。实验结果提示，ＨＤＣ前体在体外受细胞因子刺激能大量扩增。The method of isolation, purification and propagation of dendritic cell(DC) precursors from mouse liver was established domestically for the first time. The mouse livers were perfused via the portal vein using HBSS and collagenase solution and the nonparenchymal cells were separated by Percoll disontinuous density gradients centrifugation. The obtained fraction which contained DC precuresors was divided into three groups: For Group I, cells were cultured in 10% FCS RPMI 1640 supplemented with MrGM CSF and were moved onto collagen of rat tail tendon for further culture after 9 days. For Group Ⅱ, the culture medium contained MrGM CSF, but the cells were not moved. Group Ⅲ was the control group and the medium contained only 10% FCS RPMI 1640. Our results showed that after 4 days culture, many cell clusters were observed in group I and group Ⅱ, but none in group Ⅲ. After the putative liver DCs of group I were cultured one day on collagen of rat tail tendon, the clusters decreased while the DCs increased greatly in the culture media. This cell releasing phenomenon was not observed in group Ⅱ. Light and electron microscope observation showed that the purity of DC was 95%. The results indicated that hepatic DC precuresors can proliferate largely in vitro under the condition of suitable cytokines but can be mature only on collagen.

关 键 词：树突状细胞 细胞培养 细胞因子 胶原 

分 类 号：Q253[生物学—细胞生物学] Q26