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worthington 脾脏组织分离指南

发布者:欧宝体育官方 科技    发布时间:2022-06-09     
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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, 欧宝体育官方 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:脾脏组织分离指南.png

 

本技术工艺手册介绍了要求研究室步骤; 为形成受损细胞分離解决方案提拱了一大种谨守逻辑思维的研究技术; 串联出了好多聚集活性朋友参看素材,以帮到激发合理的技术。某个某个聚集的解离手册,点看到: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 / TrophoblastCollagenase Type 1: 0.1%;
Collagenase Type 2: 0.25%
DMEMHou, 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 / SpeenCollagenase Type 5: 5%;
Hyaluronidase: 2.5%;
Elastase: 1.25%
HBSSVillarreal, 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 / SplenicCollagenase Type 2: 0.1%PBSKlebanoff, 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 / SplenocytesCollagenase Type 2: 0.16%;
Deoxyribonuclease I: 0.002%
PBSYi, 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 endothelialCollagenase Type 4: 0.3-1.0%;
Deoxyribonuclease I: 20 u/ml
PBSShi, 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 / DendriticCollagenase Type 4: 0.05%RPMI 1640Abou 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 / DendriticCollagenase Type 1: 0.5%HBSSFlano, 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 / DentriticCollagenase: 300 u/ml;
Deoxyribonuclease I: 0.002%
RPMI 1640Abe, 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 / SplenicCollagenase Type 2: 0.1%;
Deoxyribonuclease I: 0.1%
RPMIMueller, 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 dendriticCollagenase: 0.1%DMEMKang, 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 stromalCollagenase Type 3: 100-400 u/mlHBSSBenedict 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 / LeukocytesCollagenase Type 4: 43 u/mlRPMI 1640Siragam, 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 / DentriticCollagenase Type 1: 0.1%;
Deoxyribonuclease I: 0.001%
RPMI-1640McLellan, 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 / DentriticCollagenase Type 3: 0.1%;
Deoxyribonuclease I: 325 u/ml
RPMI 1640Schiavoni, 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 cellsCollagenase Type 4: 0.1%;
Deoxyribonuclease I: 20 ug/ml
HEPES/RPMI 1640Stagg, 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 / DentriticCollagenase: 100 u/mlHBSSBrasel 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


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