原代细胞分离中，针对脾脏组织涉及到滋养层，脾细胞、脾脏，骨髓内皮、颊神经节，树突状和树突状细胞等等的解离通常会使用酶，如何选择合适的酶来完成对应脾脏组织的解离，以及原代细胞采集的常规操作， 艾美捷针对脾脏相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南：
本技术指南描述了标准实验室程序； 为建立细胞分离方案提供了一种合乎逻辑的实验方法； 并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。
| 脾脏 | |||||
| 种属 | 种属详情 | 细胞名称 | 酶组合推荐，含推荐浓度 | 培养基 | 参考文献 |
| 小鼠 | Mouse, 4 week | 滋养层 / Trophoblast | Collagenase Type 1: 0.1%； Collagenase Type 2: 0.25% |
DMEM | Hou, M., Han, J., Li, G., Kwon, M., Jiang, J., Emani, S., Taglauer, E., Park, J., Choi, E., Vodnala, M., Fong, Y., Emani, S., Rosas, I., Perrella, M. and Liu, X.: Multipotency of Mouse Trophoblast Stem Cells, Stem Cell Res Ther 11, 55, 2020 |
| 小鼠 | Mouse | 乳头 / Speen | Collagenase Type 5: 5%； Hyaluronidase: 2.5%； Elastase: 1.25% |
HBSS | Villarreal, E., Hewgley, W., Lang, W., Morton, C., Mao, S., Wu, J. and Sandoval, J.: In-bag Enzymatic Splenic Digestion: A Novel Alternative to Manual Morcellation?, J Surg Res 218, 209-216, 2017 |
| 小鼠 | Mouse | 脾 / Splenic | Collagenase Type 2: 0.1% | PBS | Klebanoff, C., Spencer, S., Torabi-Parizi, P., Grainger, J., Roychoudhuri, R., Ji, Y., Sukumar, M., Muranski, P., Scott, C., Hall, J., Ferreyra, G., Leonardi, A., Borman, Z. et al.: Retinoic Acid Controls the Homeostasis of Pre-cDC-Derived Splenic and Intestinal Dendritic Cells., J Exp Med 210, 1961, 2013 |
| 小鼠 | Mouse | 脾细胞 / Splenocytes | Collagenase Type 2: 0.16%； Deoxyribonuclease I: 0.002% |
PBS | Yi, T. and Cyster, J.: EBI2-Mediated Bridging Channel Positioning Supports Splenic Dendritic Cell Homeostasis and Particulate Antigen Capture., Elife 2, e00757, 2013 |
| 小鼠 | Mouse | 脾脏，骨髓内皮 / Spleen, bone marrow endothelial | Collagenase Type 4: 0.3-1.0%； Deoxyribonuclease I: 20 u/ml |
PBS | Shi, C., Jia, T., Mendez-Ferrer, S., Hohl, T., Serbina, N., Lipuma, L., Leiner, I., Li, M., Frenette, P. and Pamer, E.: Bone Marrow Mesenchymal Stem and Progenitor Cells Induce Monocyte Emigration in Response to Circulating Toll-Like Receptor Ligands., Immunity 34, 590, 2011 |
| 小鼠 | Mouse, female, 6-8 week | 树突状 / Dendritic | Collagenase Type 4: 0.05% | RPMI 1640 | Abou Fakher, F., Rachinel, N., Klimczak, M., Louis, J. and Doyen, N.: TLR9-Dependent Activation of Dendritic Cells by DNA from Leishmania Major Favors Th1 Cell Development and the Resolution of Lesions., J Immunol 182, 1386, 2009 |
| 小鼠 | Mouse | 树突状 / Dendritic | Collagenase Type 1: 0.5% | HBSS | Flano, E., Jewell, N., Durbin, R. and Durbin, J.: Methods Used to Study Respiratory Virus Infection.,Curr Protoc Cell Biol Vol. Chapter 26,, , Unit 26.3, 2009 |
| 小鼠 | Mouse | 树突状 / Dentritic | Collagenase: 300 u/ml； Deoxyribonuclease I: 0.002% |
RPMI 1640 | Abe, K., Nguyen, K., Fine, S., Mo, J., Shen, C., Shenouda, S., Corr, M., Jung, S., Lee, J., Eckmann, L. and Raz, E.: Conventional Dendritic Cells Regulate the Outcome of Colonic Inflammation Independently of T Cells., Proc Natl Acad Sci U S A 104, 17022, 2007 |
| 小鼠 | Mouse | 脾 / Splenic | Collagenase Type 2: 0.1%； Deoxyribonuclease I: 0.1% |
RPMI | Mueller, S., Matloubian, M., Clemens, D, Sharpe, A., Freeman, G., Gangappa, S., Larsen, C. and Ahmed, R.: Viral Targeting of Fibroblastic Reticular Cells Contributes to Immunosuppression and Persistence During Chronic Infection., Proc Natl Acad Sci U S A 104, 15430, 2007 |
| 小鼠 | Mouse | 脾树突状 / Splenic dendritic | Collagenase: 0.1% | DMEM | Kang, H., Liu, M. and Datta, S.: Low-dose Peptide Tolerance Therapy of Lupus Generates Plasmacytoid Dendritic Cells That Cause Expansion of Autoantigen-Specific Regulatory T Cells and Contraction of Inflammatory Th17 Cells, J Immunol 178, 7849-58, 2007 |
| 小鼠 | Mouse | 脾间质 / Splenic stromal | Collagenase Type 3: 100-400 u/ml | HBSS | Benedict Chris A, De Trez Carl, Schneider Kirsten, Ha Sukwon, Patterson Ginelle, Ware Carl F: Specific remodeling of splenic architecture by cytomegalovirus, PLoS Pathog 2, e16, 2006 |
| 小鼠 | Mouse | 白细胞 / Leukocytes | Collagenase Type 4: 43 u/ml | RPMI 1640 | Siragam, V., Crow, A., Brinc, D., Song, S., Freedman, J. and Lazarus, A.: Intravenous Immunoglobulin Ameliorates ITP via Activating Fc Gamma Receptors on Dendritic Cells., Nat Med 12, 688, 2006 |
| 小鼠 | Mouse, 4-6 week | 树突状 / Dentritic | Collagenase Type 1: 0.1%； Deoxyribonuclease I: 0.001% |
RPMI-1640 | McLellan, A., Kapp, M., Eggert, A., Linden, C., Bommhardt, U., Brocker, E., Kammerer, U. and Kampgen, E.: Anatomic Location and T-cell Stimulatory Functions of Mouse Dendritic Cell Subsets Defined by CD4 and CD8 Expression, Blood 99, 2084-93, 2002 |
| 小鼠 | Mouse | 树突状 / Dentritic | Collagenase Type 3: 0.1%； Deoxyribonuclease I: 325 u/ml |
RPMI 1640 | Schiavoni, F, Mattei, F, Sestili, P, Borghi, P, Venditti, M, Morse, H, Belardelli, F, and Gabrieli, L: ICSBP is Essential for the Development of Mouse Type I Interferon-producing Cells and for the Generation and Activation of CD8a+ Dendritic Cells, J Exp Med 196, 1415, 2002 |
| 小鼠 | Mouse | 树突状细胞 / Dentritic cells | Collagenase Type 4: 0.1%； Deoxyribonuclease I: 20 ug/ml |
HEPES/RPMI 1640 | Stagg, AJ, Burke, F, Hill, S and Knight, S.: Isolation of Mouse Spleen Dendritic Cells., Methods Mol Med 64, 9-22, 2001 |
| 小鼠 | Mouse | 树突状 / Dentritic | Collagenase: 100 u/ml | HBSS | Brasel K, De Smedt T, Smith JL, Maliszewski CR: Generation of murine dendritic cells from flt3-ligand-supplemented bone marrow cultures, Blood 96, 3029-39, 2000 |
