worthington 脾脏组织分离指南

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:

worthington 胰腺组织分离指南

 

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

关注微信公众号:每日生物评论,精彩不容错过!

每日生物评论
艾美捷科技优势代理品牌

发表评论

:?: :razz: :sad: :evil: :!: :smile: :oops: :grin: :eek: :shock: :???: :cool: :lol: :mad: :twisted: :roll: :wink: :idea: :arrow: :neutral: :cry: :mrgreen: