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Wistar大鼠树突状细胞的体外培养、扩增与鉴定
引用本文:王东海,李新钢,曲迅,杨美香,鲍修风,李刚,宫崧峰. Wistar大鼠树突状细胞的体外培养、扩增与鉴定[J]. 山东大学学报(医学版), 2006, 44(6): 568-571
作者姓名:王东海  李新钢  曲迅  杨美香  鲍修风  李刚  宫崧峰
作者单位:1. 山东大学齐鲁医院神经外科实验室,山东,济南,250012
2. 山东大学齐鲁医院临床实验中心,山东,济南,250012
摘    要:目的:探讨体外分离、培养和扩增大鼠骨髓来源树突状细胞(DC)的有效方法。方法:采用改良的Chen-WoanDC培养方法,梯度离心提取骨髓单核细胞中的非粘附细胞,加入GM CSF 5?ng/ml、IL 4 5?ng/ml进行诱导扩增,对培养的目的细胞进行形态学、OX62-FITC免疫荧光检查以及功能学鉴定。结果:DC前体细胞培养10?d呈典型树突状结构,表面标志物染色阳性,流式细胞学检查OX62单抗阳性率86%,不同培养时段的DC诱发混合淋巴细胞反应以培养10?d的DC最强,培养10?d的DC诱发混合淋巴细胞反应对同源淋巴细胞有更好的活性。结论:改良的Chen-Woan法为体外诱导扩增大鼠骨髓来源DC的有效方法。

关 键 词:树突状细胞  骨髓  细胞因子  大鼠,Wistar
文章编号:1671-7554(2006)06-0568-04
收稿时间:2005-06-21
修稿时间:2005-06-21

Induction, proliferation and identification of dendritic cells derived from rat bone marrow in vitro
WANG Dong-hai,LI Xin-gang,QU Xun,YANG Mei-xiang,BAO Xiu-feng,LI gang,GONG Song-feng. Induction, proliferation and identification of dendritic cells derived from rat bone marrow in vitro[J]. Journal of Shandong University:Health Sciences, 2006, 44(6): 568-571
Authors:WANG Dong-hai  LI Xin-gang  QU Xun  YANG Mei-xiang  BAO Xiu-feng  LI gang  GONG Song-feng
Affiliation:1. Neurosurgical Laboratory; 2. Clinical Experimental Center, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
Abstract:To study the effective method of isolation、culture and proliferation of rat bone marrow derived dendritic cells(DC) in vitro. Methods: Non adherent bone marrow monocytes were harvested by centrifugation and cultured in RPMI 1640 with cytokines( rGM CSF and rIL 4,5?ng/ml). Morphology, expression of special surface marker and functional ability were observed. Results: Morphomogical property of typical DC was observed in DC progenitors at day 10 of culture. Positive special surface marker of OX62 was observed with immunoflourense, and about 86% OX62+ cells was found by Flow cytometry analysis. To syngeneic spleen lymphocytes, the strongest proliferation of MLR induced by DC at day 10 of culture. MLR of syngeneic spleen lymphocytes was stronger than that of heterogeneous spleen lymphocytes (P<0.05). Conclusion: Improved Chen-Woan method is favorable in inducing and proliferating rat bone marrow derived DC in vitro.
Keywords:Dendritic cell   Bone marrow   Cytokine   Rat, Wistar
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