worthington 肠组织分离指南

发布者：艾美捷科技发布时间：2022-06-07

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原代细胞分离中，针对肠组织涉及到肠 L 细胞，淋巴细胞、固有层单核细胞、肠上皮和黏膜单核细胞、隐窝等等的解离通常会使用酶，如何选择合适的酶来完成对应肠组织的解离，以及原代细胞采集的常规操作， 艾美捷针对肠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：肠组织分离指南

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。

肠
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
犬/狗	Dog, adult	肠 L 细胞 / 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	淋巴细胞 / 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	微血管内皮 / 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	固有层淋巴细胞 / 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	固有层单核细胞 / 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	肠上皮 / 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	黏膜单核细胞 / 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	固有层淋巴细胞 / 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	隐窝 / 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	肠道类器官 / 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	固有层单核细胞 / 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	淋巴细胞 / 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	肠神经嵴祖细胞 / 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	结肠肿瘤类器官 / 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	肠神经元 / 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	肠道类器官 / 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	树突状，巨噬细胞 / 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	淋巴细胞 / 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	肠神经干细胞 / 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	肌间丛 / 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	固有层淋巴细胞 / 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	肠单核 / 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	肠嗜铬细胞 / 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	间质细胞 / 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	卡哈尔间质细胞 / 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