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山东大学学报(医学版)

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Wistar大鼠树突状细胞的体外培养、扩增与鉴定

王东海1,李新钢1,曲迅2,杨美香2,鲍修风1,李刚1,宫崧峰1   

  1. 山东大学齐鲁医院1. 神经外科实验室,山东 济南 250012;2. 临床实验中心,山东 济南 250012
  • 收稿日期:2006-03-29 修回日期:1900-01-01 出版日期:2006-06-24 发布日期:2006-06-24
  • 通讯作者: 王东海

WANG Dong-hai,LI Xin-gang,QU Xun,YANG Mei-xiang,BAO Xiu-feng,LI Gang,GONG Song-feng   

  • Received:2006-03-29 Revised:1900-01-01 Online:2006-06-24 Published:2006-06-24
  • Contact: WANG Dong-hai

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。

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.

Key words: Dendritic cell, Bone marrow, Cytokine, Rat, Wistar

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