Bone Marrow Derived Eosinophil Cultures
骨髓源嗜酸性细胞培养
发布: 2014年06月20日第4卷第12期 DOI: 10.21769/BioProtoc.1161 浏览次数: 14150
评审: Fanglian HeAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Feb 2013
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Abstract
Eosinophils are multifunctional effector cells implicated in the pathogenesis of a variety of diseases including asthma, eosinophil gastrointestinal disorders and helminth infection. Mouse bone marrow derived progenitor cells can be differentiated into eosinophils following IL-5 exposure. These bone marrow derived eosinophils are fully differentiated at the end of a 14 day culture based on morphology and expression of molecular markers.
Materials and Reagents
- Mice
- Histopaque 1083 (Sigma-Aldrich, catalog number: 10831 -100ml)
- IMDM with Glutamax-I (Life Technologies, InvitrogenTM, catalog number: 31980-097 )
- Fetal Bovine Serum (FBS) (Atlanta Biologicals, catalog number: S11150 )
- Penicillin/Streptomycin (Life Technologies, InvitrogenTM, catalog number: 15140-122 )
- Stem Cell Factor (Pepro tech, catalog number: 250-03 )
- FLT-3 Ligand (Pepro tech, catalog number: 250-31L )
- Recombinant mouse IL-5 (Pepro tech, catalog number: 215-15 )
- PBS (Life Technologies, InvitrogenTM, catalog number: 14200-166 )
- Diff-Quick Stain Kit (Thermo Fisher Scientific, catalog numbers: 23-122-929 , 23-122-952 , and 23-122-937 )
- RBC lysis buffer (Sigma-Aldrich, catalog number: R7757 -100ml)
- CCR3 (R&D systems, Catalog number: FAB729F )
- Siglec-F (BD, PharmingenTM, Catalog number: 552126 )
- IMDM cell culture media (see Recipes)
Equipment
- 6-well tissue culture plate
- Dissection tools: scalpel, scissors
- 1 ml syringe
- 15 ml sterile centrifuge tube
- Centrifuge
- 37 °C, 5% CO2 cell culture incubator
- Microscope
- Hemocytometer
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Lu, T. X. and Rothenberg, M. E. (2014). Bone Marrow Derived Eosinophil Cultures. Bio-protocol 4(12): e1161. DOI: 10.21769/BioProtoc.1161.
Download Citation in RIS Format
分类
免疫学 > 免疫细胞分离 > 维持和分化
细胞生物学 > 细胞分离和培养 > 细胞分化
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