小鼠骨髓来源树突状细胞（BMDC）的培养（七）
4. 培养方法的选择
本文列出迄今为止最常用的三种BMDC培养方法，我们从下表中可以更清楚地看出这三种方法的异同，相信您能根据您的需要作出正确的判断。
注：1. 该表是从文献[8]修改和添加而来；
2. 这里的Inaba法指的是其原法[2],而不是本文中的改良法。
【BMDC 培养试剂推荐】
| 生产商 | 产品名称 | 产品编号 | 产品规格 | 使用浓度 |
| PeproTech | 重组小鼠 GM-CSF | 315-03 | 5μg/20μg/50μg/100μg/250μg/500μg/1mg | 25-50ng/ml |
| PeproTech | 重组小鼠 IL-4 | 214-14 | 5μg/20μg/50μg/100μg/250μg/500μg/1mg | 10-40ng/ml |
| PeproTech | 重组小鼠 TNF-α | 315-01A | 5μg/20μg/50μg/100μg/250μg/500μg/1mg | 25-50ng/ml |
| PeproTech | 重组小鼠 sCD40L | 315-15 | 5μg/25μg/100μg/250μg/500μg/1mg | 1μg/ml |
【BMDC 鉴定试剂推荐】
| 生产商 | 产品名称 | 产品编号 | 克隆号 | 荧光标记 | 产品规格 |
| PeproTech(BioGems) | 抗小鼠 CD11c荧光标记抗体 | 3212 | N418 | FITC/PE/APC/PerCP-Cy5.5 /APC-Cy7/PE-Cy7 | 25μg/100μg/500μg |
| 抗小鼠 CD40荧光标记抗体 | 2512 | HM40-3 | FITC | 50μg/100μg/500μg/1000μg | |
| 抗小鼠 CD80荧光标记抗体 | 2912 | 16-10A1 | FITC/PE/APC | 25μg/100μg/500μg | |
| 抗小鼠 CD86荧光标记抗体 | 8912 | GL-1 | FITC/PE/APC/PE-Cy7 | 25μg/100μg/500μg | |
| 抗小鼠 MHC II类分子(I-A/I-E)荧光标记抗体 | 86212 | M5/114.15.2 | FITC/PE/APC/PE-Cy7 | 25μg/100μg/200μg /500μg |
注：用于DC鉴定的表面标志物的表达在流式图上均呈现单个峰，且多数情况下不能与阴性峰完全区分开来。为避免多色分析时补偿调节不好导致结果的不准确性，建议最好用单标，最多用双标来做DC表面标志的分析，而且必须使用同型对照，而非空白细胞对照来
排除背景染色。
【参考文献】
[1] Steinman RM, Cohn ZA.
Identification of a novel cell type in peripheral lymphoid organs of
mice. I. Morphology, quantitation, tissue distribution. J. Exp. Med.
1973; 137 （5）： 1142–62.
[2] Inaba K, Steinman RM, et. al.
Identification of proliferating dendritic cell precursors in mouse
blood. J. Exp. Med. 1992; 175（5）：1157-67.
[3] Inaba K, Inaba M,
et. al. Generation of large numbers of dendritic cells from mouse bone
marrow cultures supplemented with granulocyte/macrophage
colony-stimulating factor. J. Exp. Med. 1992; 176（6）：1693-702.
[4] Romani N, Gruner S, et. al. Proliferating dendritic cell progenitors in human blood. J. Exp.
Med. 1994; 180（1）：83-93.
[5]
Zorina T, Mayordomo JI, et. al. Culture of dendritic cells from murine
bone marrow supplemented with GM-CSF and TNF-alpha J. Immunother. 1994;
16（3）：247.
[6] Mayordomo JI, Zorina T, et. al. Bone
marrow-derived dendritic cells pulsed with synthetic tumour peptides
elicit protective and therapeutic antitumour immunity. Nat Med.
1995;1（12）：1297-302.
[7] Labeur MS, Roters B, et. al.
Generation of tumor immunity by bone marrow-derived dendritic cells
correlates with dendritic cell maturation stage. J Immunol.
1999;162（1）：168-75.
[8] Lutz MB, Kukutsch N, et. al. An advanced
culture method for generating large quantities of highly pure
dendritic cells from mouse bone marrow. J Immunol Methods.
1999;223（1）：77-92.
[9] Son YI, Egawa S, et. al. A novel
bulk-culture method for generating mature dendritic cells from mouse
bone marrow cells. J Immunol Methods. 2002; 262（1-2）：145-57.
[10]
Inaba, K., Romani, N., et al. Generation of dendritic cells from
proliferating mouse bone marrow progenitors. In: Coico, R., Ranz, A.,
Kruisbeek,A.M. （Eds.）， Current Protocols in Immunology.Wiley, New York.
1998; Unit 3.7.
