worthington 脾臟組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-06-09

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原代細(xì)胞分離中，針對(duì)脾臟組織涉及到滋養(yǎng)層，脾細(xì)胞、脾臟，骨髓內(nèi)皮、頰神經(jīng)節(jié)，樹突狀和樹突狀細(xì)胞等等的解離通常會(huì)使用酶，如何選擇合適的酶來完成對(duì)應(yīng)脾臟組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對(duì)脾臟相關(guān)的組織分類解離分種屬、不同的細(xì)胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻(xiàn)，詳細(xì)匯總指南：脾臟組織分離指南.png

本技術(shù)指南描述了標(biāo)準(zhǔn)實(shí)驗(yàn)室程序；為建立細(xì)胞分離方案提供了一種合乎邏輯的實(shí)驗(yàn)方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點(diǎn)擊查看：各組織分離指南。

脾臟
種屬	種屬詳情	細(xì)胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻(xiàn)
小鼠	Mouse, 4 week	滋養(yǎng)層 / 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	乳頭 / 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	脾細(xì)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	脾臟，骨髓內(nèi)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	樹突狀 / 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	樹突狀 / 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	樹突狀 / 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	脾樹突狀 / 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	脾間質(zhì) / 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	白細(xì)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	樹突狀 / 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	樹突狀 / 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	樹突狀細(xì)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	樹突狀 / 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