大鼠骨髓树突状细胞的分离和培养  

Isolation,culture and immunological character of DC from F344 rat bone marrow

作　　者：杨贺勤[1] 张震宇[2] 

机构地区：[1]首都医科大学宣武医院妇产科,北京100053 [2]首都医科大学附属北京朝阳医院妇产科

出　　处：《现代妇产科进展》2009年第7期486-489,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目(No:39970770)

摘　　要：目的:探讨体外分离和培养大鼠源性树突状细胞的方法,及其经卵巢肿瘤提取物致敏后在体内诱发抗肿瘤免疫效应。方法:(1)取大鼠骨髓悬液,经Tris-NH4Cl裂解红细胞后得到大鼠骨髓单个核细胞;(2)应用大鼠重组粒细胞-巨噬细胞集落刺激因子(rrGM-CSF)、大鼠重组白细胞介素4(rrIL-4)、大鼠重组肿瘤坏死因子-α(rrTNF-α)进行体外培养;(3)培养第5天时加入NuTu-19 Fischer 344大鼠卵巢肿瘤细胞提取物,获得负载卵巢肿瘤提取物的树突状细胞;(4)体内实验检测DC诱发抗肿瘤免疫效力。结果:(1)大鼠骨髓来源的骨髓单个核细胞(bone marrow mononuclear cells,BMMNC)在相关细胞因子作用下可培养出成熟的树突状细胞;(2)体内实验表明,预先免疫的大鼠肿瘤出现时间较晚,肿瘤生长速度较慢,与对照组有显著差异(P<0.05),抑瘤率59.4%;治疗组大鼠在实验过程中肿瘤体积小,最终肿瘤质量轻,与对照组有显著差异(P<0.05),与先行免疫组无明显差异(P>0.05),抑瘤率69.3%。结论:大鼠骨髓来源的BMMNC可培养出成熟的树突状细胞。DC可负载并提呈肿瘤提取物,体内实验显示肿瘤提取物致敏的树突状细胞可以杀伤肿瘤细胞。Objective：To explore the methods for isolating and culturing dendritic cells （DCs） derived from F344 rat bone marrow,and the anti-tumor response in vivo induced by DCs pulsed by ovarian tumor extracts. Methods： （1） The bone marrow mononuclear cells （BMMNCs） were obtained from bone marrow of F344 rats＇limbs through spalled RBC by Tris-NH4C1. （2）BMMNCs were cultured in medium with granulocyte-macrophage colony stimulating factor （ GM-CSF）, interleukin-4 （IL-4） and tumor necrosis factor-or （TNF-α）. （ 3 ） After culturing for 5 days, added the rats ovarian cancer cell line NuTu-19 extracts to pulse DC and acquired the extracts pulsed DCs. （4）The anti-tumor response induced by DCs pulsed with ovarian tumor extracts were analyzed in vivo. Results： （ 1 ） The mature DC could be induced by BMMNC cocultured with relative cytokines. （2）In vivo, the pre-immune group had a slower tumor growth rate and occurring tumor later, which exists obvious difference from the control group（ P 〈 0.05 ）, and its inhibition ratio was 59.4%. During the experience, tumor volume and tumor weight were reduced in rats vaccinated with DC, compared with rats vaccinated with PBS （ P 〈 0.05 ） and pre-immune group （ P 〉 0.05 ）, and its inhibition rate was 69.3%. Conclusion：The mature DC can be obtained from the BMMNC. DC loaded tumor extracts can induce active immuity against tumor cells in vivo.

关 键 词：树突状细胞 大鼠 近交F344 肿瘤提取物 细胞培养 

分 类 号：R737.33[医药卫生—肿瘤]