worthington 腸組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-06-07

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原代細(xì)胞分離中，針對(duì)腸組織涉及到腸 L 細(xì)胞，淋巴細(xì)胞、固有層單核細(xì)胞、腸上皮和黏膜單核細(xì)胞、隱窩等等的解離通常會(huì)使用酶，如何選擇合適的酶來完成對(duì)應(yīng)腸組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對(duì)腸相關(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：腸組織分離指南

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。

腸
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)
犬/狗	Dog, adult	腸 L 細(xì)胞 / Intestinal L-cells	Collagenase Type 1: 75 u/ml	HBSS	Damholt AB, Buchan AM, Kofod H: Glucagon-like-peptide-1 secretion from canine L-cells is increased by glucose-dependent-insulinotropic peptide but unaffected by glucose, Endocrinology 139, 2085-91, 1998
人	Human	淋巴細(xì)胞 / Lymphocytes	CLSPA: 0.05%； Deoxyribonuclease I: 0.00075%	RPMI-1640	Trapecar, M., Khan, S., Roan, N., Chen, T., Telwatte, S., Deswal, M., Pao, M., Somsouk, M., Deeks, S., Hunt, P., Yukl, S. and Sanjabi, S.: An Optimized and Validated Method for Isolation and Characterization of Lymphocytes from HIV+ Human Gut Biopsies., AIDS Res Hum Retroviruses 33, S31-S39, 2017
人	Human	微血管內(nèi)皮 / Microvessel endothelial	Collagenase Type 2: 0.2%	HBSS	Kessler, S., Rho, H., West, G., Fiocchi, C., Drazba, J. and de la Motte, C.: Hyaluronan (HA) Deposition Precedes and Promotes Leukocyte Recruitment in Intestinal Inflammation., Clin Transl Sci 1, 57-61, 2008
人	Human	固有層淋巴細(xì)胞 / Lamina propria lymphocytes	Collagenase: 20 u/ml	Medium	Ebert, Ellen.: CD2 Activation of Human Lamina Propria Lymphocytes Reduces CD3 Responsiveness., Immunology 117, 71-7, 2006
人	Human	固有層單核細(xì)胞 / Lamina propria mononuclear cells	Collagenase: 0.02%	HBSS	Kanai Takanori, Totsuka Teruji, Uraushihara Koji, Makita Shin, Nakamura Tetsuya, Koganei Kazutaka, Fukushima Tsuneo, Akiba Hisaya, Yagita Hideo, Okumura Ko, Machida Utako, Iwai Hideyuki, Azuma Miyuki, Chen Lieping, Watanabe Mamoru: Blockade of B7-H1 suppresses the development of chronic intestinal inflammation, J Immunol 171, 4156-63, 2003
人	Human	腸上皮 / Intestinal epithelial	Collagenase Type 4: 72.5 u/ml	HBSS	Fahlgren, A., Hammarstrom, S., Danielsson, A. and Hammarstrom, M.: Increased Expression of Antimicrobial Peptides and Lysozyme in Colonic Epithelial Cells of Patients with Ulcerative Colitis., Clin Exp Immunol 131, 90, 2003
人	Human	黏膜單核細(xì)胞 / Mucosal mononuclear cells	Collagenase Type 3: 0.01%； Deoxyribonuclease I: 0.01%； Soybean Trypsin Inhibitor: 0.01%	RPMI	Stallmach A, Schaffer F, Hoffmann S, Weber S, Muller-Molaian I, Schneider T, Kohne G, Ecker KW, Feifel G, Zeitz M: Increased state of activation of CD4 positive T cells and elevated interferon gamma production in pouchitis, Gut 43, 499-505, 1998
小鼠	Mouse	固有層淋巴細(xì)胞 / Lamina propria lymphocytes	Collagenase Type 4: 0.02%； Deoxyribonuclease I: 0.01%	HBSS	Emgard, J., Kammoun, H., Garcia-Cassani, B., Chesne, J., Parigi, S., Jacob, J., Cheng, H, Evren, E., Das, S., Czarnewski, P., Sleiers, N. Melo-Gonzalez, F., Kvedaraite, E. Svensson, M. and Scandella, E.: Oxysterol Sensing through the Receptor GPR183 Promotes the Lymphoid-Tissue-Inducing Function of Innate Lymphoid Cells and Colonic Inflammation., Immunity 48, 120-132.e8, 2018
小鼠	Mouse, embryonic	隱窩 / Crypt	Collagenase Type 1: 200 u/ml； Hyaluronidase: 100u/ml	DMEM	Ren HJ, Zhang CL, Liu RD, Li N, Li XG, Xue HK, Guo Y, Wang ZQ, Cui J and Ming L: Primary Cultures of Mouse Small Intestinal Epithelial Cells using the Dissociating Enzyme Type I Collagenase and Hyaluronidase., Braz J Med Biol Res 50, e5831, 2017
小鼠	Mouse	腸道類器官 / Intestinal organoids	Collagenase Type 4: 500 u/ml； Deoxyribonuclease I: 15 u/ml	Basal Medium	O'Rourke, K., Ackerman, S., Dow, L. and Lowe, S.: Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells., Bio Protoc 6, , 2016
小鼠	Mouse	固有層單核細(xì)胞 / Lamina propria mononuclear cells	Collagenase Type 3: 400 u/ml； Deoxyribonuclease I: 0.001%	HBSS	Seo, S., Kuffa, P., Kitamoto, S., Nagao-Kitamoto, H., Rousseau, J., Kim, Y., Nunez, G. and Kamada, N.: Intestinal Macrophages Arising from CCR2(+) Monocytes Control Pathogen Infection by Activating Innate Lymphoid Cells., Nat Commun 6, 8010, 2015
小鼠	Mouse	淋巴細(xì)胞 / Lymphocytes	Collagenase Type 1: 100 u/ml	RPMI-1640	Beura, L., Anderson, K., Schenkel, J., Locquiao, J., Fraser, K., Vezys, V., Pepper, M. and Masopust, D.: Lymphocytic Choriomeningitis Virus Persistence Promotes Effector-Like Memory Differentiation and Enhances Mucosal T cell Distribution., J Leukoc Biol 97, 217, 2015
小鼠	Mouse, P4	腸神經(jīng)嵴祖細(xì)胞 / Enteric neural crest progenitors	Neutral Protease: 0.5%； Collagenase animal free: 0.05%	DMEM/F12	Hotta, R., Stamp, L., Foong, J., McConnell, S., Bergner, A., Anderson, R., Enomoto, H., Newgreen, D., Obermayr, F., Furness, J. and Young, H.: Transplanted Progenitors Generate Functional Enteric Neurons in the Postnatal Colon., J Clin Invest 123, 1182, 2013
小鼠	Mouse	結(jié)腸腫瘤類器官 / Colon tumor organoids	Collagenase Type 4: 200 u/ml； Neutral Protease: 0.01%	DMEM	Xue, X. and Shah, Y.: In Vitro Organoid Culture of Primary Mouse Colon Tumors., J Vis Exp , e50210, 2013
小鼠	Mouse	腸神經(jīng)元 / Enteric neurons	Collagenase Type 4: 0.1%； Trypsin: 0.05%； Deoxyribonuclease I: 400 u/ml	DMEM/F12	Zhang, Y. and Hu, W.: Mouse Enteric Neuronal Cell Culture., Methods Mol Biol 1078, 55-63, 2013
小鼠	Mouse	腸道類器官 / Intestinal organoids	Collagenase Type 1: 800 u/ml； Neutral Protease: 0.013%	DMEM	Barthel, E., Speer, A., Levin, D., Sala, F., Hou, X., Torashima, Y., Wigfall, C. and Grikscheit, T.: Tissue Engineering of the Intestine in a Murine Model., J Vis Exp 70, e4279, 2012
小鼠	Mouse	樹突狀，巨噬細(xì)胞 / Dendritic, macrophages	Collagenase: 0.15%； Deoxyribonuclease I: 0.004%	HBSS	Geem, D., Medina-Contreras, O., Kim, W., Huang, C. and Denning T.: Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine., J Vis Exp 63, e4040, 2012
小鼠	Mouse	淋巴細(xì)胞 / Lyphocytes	Collagenase Type 4: 0.05%； Neutral Protease: 0.1%； Deoxyribonuclease I: 200 ug/ml	RPMI 1640	Ruiz, V., Sachdev, M., Zhang, S., Wen, S. and Moss, S.: Isolating, Immunophenotyping and Ex vivo Stimulation of CD4+ and CD8+ Gastric Lymphocytes During Murine Helicobacter Pylori Infection., J Immunol Methods 384, 157-63, 2012
小鼠	Mouse, 2-3 month	腸神經(jīng)干細(xì)胞 / Enteric neural stem cells	Collagenase: 0.11%； Neutral Protease: 0.1%； Deoxyribonuclease I: 50 u/ml	M199	Becker, L., Kulkarni, S., Tiwari, G., Micci, M. and Pasricha, P.: Divergent Fate and Origin of Neurosphere-Like Bodies from Different Layers of the Gut., Am J Physiol Gastrointest Liver Physiol 302, G958-65, 2012
小鼠	Mouse, neonatal and adult	肌間叢 / Myenteric plexus	Collagenase Type 4: 0.025-0.1%； Papain: 10 u/ml； Deoxyribonuclease I: 100 u/ml	HBSS	Joseph, N., He, S., Quintana, E., Kim, Y., Nunez, G. and Morrison, S.: Enteric Glia are Multipotent in Culture but Primarily Form Glia in the Adult Rodent Gut., J Clin Invest 121, 3398, 2011
小鼠	Mouse	固有層淋巴細(xì)胞 / Lamina propria lymphocytes	Collagenase Type 1: 0.3%； Deoxyribonuclease I: 0.01%	RPMI 1640	Ito, Y., Kanai, T., Totsuka, T., Okamoto, R., Tsuchiya, K., Nemoto, Y., Yoshioka, A., Tomita, T., Nagaishi, T., Sakamoto, N., Sakanishi, T., Okumura, K., Yagita, H. and Watanabe, M.: Blockade of NKG2D Signaling Prevents the Development of Murine CD4+ T Cell-Mediated Colitis., Am J Physiol Gastrointest Liver Physiol 294, G199, 2008
小鼠	Mouse	腸單核 / Intestinal mononuclear	Collagenase Type 4: 0.1%	RPMI 1640	Forbes, E., Groschwitz, K., Abonia, J.P., Brandt, E., Cohen, E., Blanchard, C., Ahrens, R., Seidu, L., McKenzie, A., Strait, R., Finkelman, F., Foster, P., Matthaei, K., Rothenberg, M. and Hogan, S.: IL-9- and Mast Cell-Mediated Intestinal Permeability Predisposes to Oral Antigen Hypersensitivity., J Exp Med 205, 897, 2008
小鼠	Mouse, Mastomys natalensis	腸嗜鉻細(xì)胞 / Enterochromaffin cells	Collagenase: 0.025%； Pronase E: 0.07%	HBSS	Kidd, M., Modlin, I., Eick, G. and Champaneria, M.: Isolation, Functional Characterization, and Transcriptome of Mastomys Ileal Enterochromaffin Cells., Am J Physiol Gastrointest Liver Physiol 291, G778-91, 2006
小鼠	Mouse, 7 week	間質(zhì)細(xì)胞 / Interstitial cells	Collagenase: 0.04%； Trypsin: 0.02%	see reference	Goto Kazunori, Matsuoka Satoshi, Noma Akinori: Two types of spontaneous depolarizations in the interstitial cells freshly prepared from the murine small intestine, J Physiol 559, 411-22, 2004
小鼠	Mouse, 9-13 day	卡哈爾間質(zhì)細(xì)胞 / Interstitial cells of Cajal	Collagenase Type 2: 0.13%	HBSS	Ordag Tamas, Redelman Doug, Miller Lisa J, Horvath Viktor J, Zhong Qiao, Almeida-Porada Graca, Zanjani Esmail D, Horowitz Burton, Sanders Kenton M: Purification of interstitial cells of Cajal by fluorescence-activated cell sorting, Am J Physiol Cell Physiol 286, C448-56, 2004