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worthington 结肠组织分离指南

发布者:酷游ku119网址 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, 酷游ku119网址 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 / EnterochromaffinTrypsin: 0.05%;
Collagenase Type 1: 0.1%
DMEMRaghupathi, 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 gangliaCollagenase Type 4: 0.2%;
Protease: 0.1%
Kreb's solutionKang, 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
Human固有层单核 / Lamina propria mononuclearCollagenase Type 1: 0.1%;
Deoxyribonuclease I: 5 u/ml
HBSSMoser, 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
Human结肠肠神经 / Colon enteric neuralNeutral Protease: 0.5%;
Collagenase animal free: 0.1%;
Hyaluronidase: 0.1%;
Deoxyribonuclease I: 0.01%
Ham's F-12Rollo, 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
Human胃肠上皮 / Gastrointestinal epithelialCollagenase Type 1: 0.2%DMEM/F12VanDussen, 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
Human结肠 / ColonTrypsin: 0.1%;
Collagenase: 0.1%
HBSSAli, 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
Human肠嗜铬 / EnterochromaffinTrypsin: 0.05%;
Collagenase Type 1: 0.1%
DMEMRaghupathi, 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
Human结肠上皮 / Colonic epithelialCollagenase: 150 u/ml;
Neutral Protease: 0.04 mg/ml
Basal X mediaRoig, 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
Human大肠癌 / Colorectal cancerCollagenase Type 4: 1%;
Deoxyribonuclease I: 0.2%
HBSSZhou, 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
Human结肠上皮 / Colonic epithelialCollagenase Type 4: 0.1%not listedHuang, 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
Human结肠癌 / Colon cancerCollagenase Type 1: 300 u/ml;
Hyaluronidase: 100 u/ml
DMEM/F12Varnat, 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
Human癌症干细胞 / Cancer stem cellCollagenase Type 3: 200 u/ml;
Deoxyribonuclease I: 100 u/ml
RPMI-1640Dalerba 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
Human结肠上皮 / Colonic epithelialCollagenase:;
Neutral Protease: 0.3%;
Deoxyribonuclease I: 0.05%
RPMI 1640Fukushima, 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
Human结肠内皮细胞 / Colonic endothelial cellsCollagenase Type 2: 0.25%HBSS/5%FBSWang 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
Human结肠细胞 / ColonocytesCollagenase: 0.15%DMEM/F12Emenaker 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
Human上皮和黏膜淋巴细胞 / Epithelial and mucosal lymphocytesNeutral Protease: 0.1%;
CLSPA: 0.02%;
Deoxyribonuclease I: 0.01%
RPMI 1640Hisamatsu, 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
Human结肠上皮 / Colonic epithelialNeutral Protease: 1.2 u/ml;
Collagenase Type 4: 50 u/ml
HBSSGibson, 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
Human固有层淋巴细胞 / Lamina propria lymphocytesCollagenase: 25 u/mlHBSSUeyama 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 propriaCollagenase: 100 u/ml;
Deoxyribonuclease I: 0.004%
RPMI 1640Larmonier, 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 organoidCollagenase Type 1: 800 u/ml;
Neutral Protease: 0.013%
DMEMFattahi, 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 lymphocytesNeutral Protease: 3 u/ml;
Collagenase Type 4: 0.05%
HBSSRound, 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 propriaCollagenase Type 2: 0.1%;
Neutral Protease: 0.1%;
Deoxyribonuclease I: 0.004%
RPMI 1640Atarashi, 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 cellCollagenase Type 3: 200 u/ml;
Deoxyribonuclease I: 100 u/ml
RPMI-1640Dalerba 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 cellsCollagenase: 0.05%;
Deoxyribonuclease I: 0.05%;
Neutral Protease: 0.3%
HBSSWeigmann 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 / 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淋巴细胞 / LympocytesCollagenase/Dispase: 100 u/mlRPMI 1640Annacker 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 lymphocytesCollagenase Type 1: 0.2%;
Deoxyribonuclease I: 0.01%
HBSSTotsuka 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 cellsCollagenase Type 2: 0.015%;
Deoxyribonuclease I: 0.01%
RPMIWirtz 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


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