worthington 结肠组织分离指南

发布者：艾美捷科技发布时间：2022-06-01

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原代细胞分离中，针对结肠组织涉及到肠嗜铬、肌间神经节、结肠和结肠上皮、结肠上皮等等的解离通常会使用酶，如何选择合适的酶来完成对应结肠组织的解离，以及原代细胞采集的常规操作， 艾美捷针对结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：

结肠组织分离指南

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。

结肠
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
豚鼠	Guinea-pig, adult	肠嗜铬 / Enterochromaffin	Trypsin: 0.05%； Collagenase Type 1: 0.1%	DMEM	Raghupathi, R., Duffield, M., Zelkas, L., Meedeniya, A., Brookes, S., Sia, T., Wattchow, D., Spencer, N. and Keating, D.: Identification of Unique Release Kinetics of Serotonin From Guinea-pig and Human Enterochromaffin Cells., J Physiol 591, 5959-75, 2013
豚鼠	Guinea-Pig, male, 200-250g	肌间神经节 / Myenteric ganglia	Collagenase Type 4: 0.2%； Protease: 0.1%	Kreb's solution	Kang, M., Maguma, H., Smith, T., Ross, G., Dewey, W. and Akbarali, H.: The Role of β-arrestin2 in the Mechanism of Morphine Tolerance in the Mouse and Guinea Pig Gastrointestinal Tract., J Pharmacol Exp Ther 340, 567-76, 2012
人	Human	固有层单核 / Lamina propria mononuclear	Collagenase Type 1: 0.1%； Deoxyribonuclease I: 5 u/ml	HBSS	Moser, A., Spindelboeck, W., Strohmaier, H., Enzinger, C., Gattringer, T., Fuchs, S., Fazekas, F., Gorkiewicz, G., Wurm, P., Hogenauer, C. and Khalil, M.: Mucosal Biopsy Shows Immunologic Changes of the Colon in Patients with Early MS, Neurol Neuroimmunol Neuroinflamm 4, e362, 2017
人	Human	结肠肠神经 / Colon enteric neural	Neutral Protease: 0.5%； Collagenase animal free: 0.1%； Hyaluronidase: 0.1%； Deoxyribonuclease I: 0.01%	Ham's F-12	Rollo, B., Zhang, D., Stamp, L., Menheniott, T., Stathopoulos, L., Denham, M., Dottori, M., King, S., Hutson, J. and Newgreen, D.: Enteric Neural Cells from Hirschsprung Disease Patients Form Ganglia in Autologous Aneuronal Colon, Cell Mol Gastroenterol Hepatol 2, 92, 2016
人	Human	胃肠上皮 / Gastrointestinal epithelial	Collagenase Type 1: 0.2%	DMEM/F12	VanDussen, K., Marinshaw, J., Shaikh, N., Miyoshi, H., Moon, C., Tarr, P., Ciorba, M. and Stappenbeck, T.: Development of an Enhanced Human Gastrointestinal Epithelial Culture System to Facilitate Patient-Based Assays., Gut 64, 911-20, 2015
人	Human	结肠 / Colon	Trypsin: 0.1%； Collagenase: 0.1%	HBSS	Ali, M., Anand, S., Tangella, K., Ramkumar, D. and Saif, T.: Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry., J Vis Exp , e52532, 2015
人	Human	肠嗜铬 / Enterochromaffin	Trypsin: 0.05%； Collagenase Type 1: 0.1%	DMEM	Raghupathi, R., Duffield, M., Zelkas, L., Meedeniya, A., Brookes, S., Sia, T., Wattchow, D., Spencer, N. and Keating, D.: Identification of Unique Release Kinetics of Serotonin From Guinea-pig and Human Enterochromaffin Cells., J Physiol 591, 5959-75, 2013
人	Human	结肠上皮 / Colonic epithelial	Collagenase: 150 u/ml； Neutral Protease: 0.04 mg/ml	Basal X media	Roig, A., Eskiocak, U., Hight, S., Kim, S., Delgado, O., Souza, R., Spechler, S., Wright, W. and Shay, J.: Immortalized Epithelial Cells Derived from Human Colon Biopsies Express Stem Cell Markers and Differentiate In Vitro., Gastroenterol 138, 1012, 2010
人	Human	大肠癌 / Colorectal cancer	Collagenase Type 4: 1%； Deoxyribonuclease I: 0.2%	HBSS	Zhou, J., Belov, L., Huang, P., Shin, J., Solomon, M., Chapuis, P., Bokey, L., Chan, C., Clarke, C., Clarke, S. and Christopherson, R.: Surface Antigen Profiling of Colorectal Cancer Using Antibody Microarrays With Fluorescence Multiplexing., J Immunol Methods 355, 40, 2010
人	Human	结肠上皮 / Colonic epithelial	Collagenase Type 4: 0.1%	not listed	Huang, E., Hynes, M., Zhang, T., Ginestier, C., Dontu, G., Appelman, H., Fields, J., Wicha, M. and Boman, B.: Aldehyde Dehydrogenase 1 is a Marker for Normal and Malignant Human Colonic Stem Cells (SC) and Tracks SC Overpopulation During Colon Tumorigenesis., Cancer Res 69, 3382-9, 2009
人	Human	结肠癌 / Colon cancer	Collagenase Type 1: 300 u/ml； Hyaluronidase: 100 u/ml	DMEM/F12	Varnat, F., Duquet, A., Malerba, M., Zbinden, M., Mas, C., Gervaz, P. and Ruiz i Altaba, A.: Human Colon Cancer Epithelial Cells Harbour Active HEDGEHOG-GLI Signalling that is Essential for Tumour Growth, Recurrence, Metastasis and Stem Cell Survival and Expansion., EMBO Mol Med 1, 338-51, 2009
人	Human	癌症干细胞 / Cancer stem cell	Collagenase Type 3: 200 u/ml； Deoxyribonuclease I: 100 u/ml	RPMI-1640	Dalerba Piero, Dylla Scott J, Park In-Kyung, Liu Rui, Wang Xinhao, Cho Robert W, Hoey Timothy, Gurney Austin, Huang Emina H, Simeone Diane M, Shelton Andrew A, Parmiani Giorgio, Castelli Chiara, Clarke Michael F: Phenotypic characterization of human colorectal cancer stem cells, Proc Natl Acad Sci U S A 104, 10158-63, 2007
人	Human	结肠上皮 / Colonic epithelial	Collagenase:； Neutral Protease: 0.3%； Deoxyribonuclease I: 0.05%	RPMI 1640	Fukushima, K. and Fiocchi, C.: Paradoxical Decrease of Mitochondrial DNA Deletions in Epithelial Cells of Active Ulcerative Colitis Patients., Am J Physiol Gastrointest Liver Physiol 286, G804-13, 2004
人	Human	结肠内皮细胞 / Colonic endothelial cells	Collagenase Type 2: 0.25%	HBSS/5%FBS	Wang D., Lehman R., Donner D., Matli M., Warren R., and Welton M.: Expression and Endocytosis of VEGF and Its Receptors in Human Colonic Vascular Endothelial Cells, Am J Physiol/Gastro 282, G1088, 2002
人	Human	结肠细胞 / Colonocytes	Collagenase: 0.15%	DMEM/F12	Emenaker N, Calaf G, Cox D, Basson M and Qureshi N: Short Chain Fatty Acids Differentially Modulate Cellular Phenotype and c-myc Protein Levels in Primary Human Nonmalignant and Malignant Colonocytes, J Nutr 46, 96-105, 2001
人	Human	上皮和黏膜淋巴细胞 / Epithelial and mucosal lymphocytes	Neutral Protease: 0.1%； CLSPA: 0.02%； Deoxyribonuclease I: 0.01%	RPMI 1640	Hisamatsu, T., Watanabe, M., Ogata, H., Ezaki, T., Hozawa, S., Ishii, H., Kanai, T. and Hibi, T.: Interferon-Inducible Gene Family 1-8U Expression in Colitis-Associated Colon Cancer and Severely Inflamed Mucosa in Ulcerative Colitis., Cancer Res 59, 5927-31, 1999
人	Human	结肠上皮 / Colonic epithelial	Neutral Protease: 1.2 u/ml； Collagenase Type 4: 50 u/ml	HBSS	Gibson, P., Rosella, O., Wilson, A., Mariadason, J., Rickard, K., Byron, K. and Barkla, D.: Colonic Epithelial Cell Activation and the Paradoxical Effects of Butyrate., Carcinogenesis 20, 539, 1999
人	Human	固有层淋巴细胞 / Lamina propria lymphocytes	Collagenase: 25 u/ml	HBSS	Ueyama H, Kiyohara T, Sawada N, Isozaki K, Kitamura S, Kondo S, Miyagawa J, Kanayama S, Shinomura Y, Ishikawa H, Ohtani T, Nezu R, Nagata S, Matsuzawa Y: High Fas ligand expression on lymphocytes in lesions of ulcerative colitis, Gut 43, 48-55, 1998
小鼠	Mouse	固有层 / Lamina propria	Collagenase: 100 u/ml； Deoxyribonuclease I: 0.004%	RPMI 1640	Larmonier, C., Shehab, K., Laubitz, D., Jamwal, D., Ghishan, F. and Kiela, P.: Transcriptional Reprogramming and Resistance to Colonic Mucosal Injury in Poly(ADP-ribose) Polymerase 1 (PARP1)-deficient Mice., J Biol Chem 291, 8918-30, 2016
小鼠	Mouse	结肠类器官 / Colon organoid	Collagenase Type 1: 800 u/ml； Neutral Protease: 0.013%	DMEM	Fattahi, F., Steinbeck, J., Kriks, S., Tchieu, J., Zimmer, B., Kishinevsky, S., Zeltner, N., Mica, Y., El-Nachef, W., Zhao, H., de Stanchina, E., Gershon, M., Grikscheit, T., Chen, S. and Studer, L.: Deriving Human ENS Lineages for Cell Therapy and Drug Discovery in Hirschsprung Disease., Nature 531, 105-9, 2016
小鼠	Mouse, 8-10 week	固有层淋巴细胞 / Lamina propria lymphocytes	Neutral Protease: 3 u/ml； Collagenase Type 4: 0.05%	HBSS	Round, J. and Mazmanian, S.: Inducible Foxp3+ Regulatory T-cell Development by a Commensal Bacterium of the Intestinal Microbiota., Proc Natl Acad Sci U S A 107, 12204-9, 2010
小鼠	Mouse	固有层 / Lamina propria	Collagenase Type 2: 0.1%； Neutral Protease: 0.1%； Deoxyribonuclease I: 0.004%	RPMI 1640	Atarashi, K., Nishimura, J., Shima, T., Umesaki, Y., Yamamoto, M., Onoue, M., Yagita, H., Ishii, N., Evans, R., Honda, K. and Takeda, K.: ATP Drives Lamina Propria T(H)17 Cell Differentiation., Nature 455, 808, 2008
小鼠	Mouse, 6-8 week	癌症干细胞 / Cancer stem cell	Collagenase Type 3: 200 u/ml； Deoxyribonuclease I: 100 u/ml	RPMI-1640	Dalerba Piero, Dylla Scott J, Park In-Kyung, Liu Rui, Wang Xinhao, Cho Robert W, Hoey Timothy, Gurney Austin, Huang Emina H, Simeone Diane M, Shelton Andrew A, Parmiani Giorgio, Castelli Chiara, Clarke Michael F: Phenotypic characterization of human colorectal cancer stem cells, Proc Natl Acad Sci U S A 104, 10158-63, 2007
小鼠	Mouse	固有层单核细胞 / Lamina propria mononuclear cells	Collagenase: 0.05%； Deoxyribonuclease I: 0.05%； Neutral Protease: 0.3%	HBSS	Weigmann Benno, Tubbe Ingrid, Seidel Daniel, Nicolaev Alex, Becker Christoph, Neurath Markus F: Isolation and subsequent analysis of murine lamina propria mononuclear cells from colonic tissue, Nat Protoc 2, 2307-11, 2007
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	淋巴细胞 / Lympocytes	Collagenase/Dispase: 100 u/ml	RPMI 1640	Annacker O, Coombes JL, Malmstrom V, Uhlig HH, Bourne T, Johansson-Lindbom B, Agace WW, Parker CM, Powrie F.: Essential role for CD103 in the T cell-mediated regulation of experimental colitis, J Exp Med 202, 1051, 2005
小鼠	Mouse, 6-8 week	固有层淋巴细胞 / Lamina propria lymphocytes	Collagenase Type 1: 0.2%； Deoxyribonuclease I: 0.01%	HBSS	Totsuka T, Kanai T, Uraushihara K, Iiyama R, Yamazaki M, Akiba H, Yagita H, Okumura K, Watanabe M: Therapeutic effect of anti-OX40L and anti-TNF-alpha MAbs in a murine model of chronic colitis, Am J Physiol/Gastro 284, G595-603, 2003
小鼠	Mouse, 6-8 week	固有层单核细胞 / Lamina propria mononuclear cells	Collagenase Type 2: 0.015%； Deoxyribonuclease I: 0.01%	RPMI	Wirtz S., Becker C., Blumberg R., Galle P., and Neurath M.: Treatment of T cell-dependent experimental colitis in SCID mice by local administration of an adenovirus expressing IL-18 antisense mRNA, J Immunol 168(1), 411, 2002