Worthington 肺组织分离指南

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，针对肺细胞组织涉及到内皮，肺、肺泡Ⅱ型、腺泡，肺肿瘤，肺泡上皮和成纤维细胞等等的解离通常会使用酶，如何选择合适的酶来完成对应肺细胞组织的解离，以及原代细胞采集的常规操作， 艾美捷针对肺细胞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：肺组织分离指南

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。

肺
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
牛	Bovine	内皮，肺 / Endothelial，Pulmonary	Collagenase: 1000 u/ml； Collagenase Type 2: 1000 u/ml	PBS, CMF	Del Vecchio, P., Siflinger-Birnboim, A., Belloni, P., Holleran, L., Lum, H., and Malik, A.: Culture and Characterization of Pulmonary Microvascular Endothelial Cells, In Vitro Cell Dev Biol 28A, 711, 1992
豚鼠	Guinea-pig, male, 250-300 g	肺泡Ⅱ型 / Alveolar type II	Elastase: 40 u/ml	PBS (see reference)	Sikpi, M., Nair, C., Johns, A., and Das, S.: Metabolic and Ultrastructural Characterizationof Guinea-Pig Alveolar Type II Cells Isolated by Centrifugal Elutriation, Biochim Biophys Acta 877, 20, 1986
豚鼠	Guinea-pig, male, 100 g	单细胞 / Single cells	Trypsin: 0.25%	CF salt solution	Phillips, H.: Dissociation of Single Cells from Lung or Kidney Tissue with Elastase, In Vitro 8, 101, 1972
人	Human	腺泡 / Aveolar	Elastase: 12.9 u/ml	PBS	Bahmed, K., Boukhenouna, S., Karim, L., Andrews, T., Lin, J., Powers, R., Wilson, M., Lin, C., Messier, E., Reisdorph, N., Powell, R., Tang, H., Mason, R., Criner, G. and Kosmider, B.: The Effect of Cysteine Oxidation on DJ-1 Cytoprotective Function in Human Alveolar Type II Cells., Cell Death Dis 10, 638, 2019
人	Human	肺肿瘤 / Lung tumor	Collagenase Type 1: 45-60 u/ml； Collagenase Type 2: 15-20 u/ml； Collagenase Type 4: 45-60 u/ml； Elastase: 0.002%； Deoxyribonuclease I: 0.002%	DMEM/F12	Quatromoni, J., Singhal, S., Bhojnagarwala, P., Hancock, W., Albelda, S. and Eruslanov, E.: An Optimized Dissagregation Method for Human Lung Tumors That Perserves the Phenotype and Function of the Immune Cells, J Leukoc Biol 97, 201, 2015
人	Human	肺泡上皮 / Alveolar epithelial	Neutral Protease: 2 u/ml； Trypsin: 0.25%； Elastase: 10 u/ml	Bronchial Epithelial Growth Medium	Barkauskas, C., Cronce, M., Rackley, C., Bowie, E., Keene, D., Stripp, B., Randell, S., Noble, P. and Hogan, B.: Type 2 Alveolar Cells are Stem Cells in Adult Lung, J Clin Invest 123, 3025, 2013
人	Human	肺内皮 / Pulmonary endothelial	Collagenase Type 2: 0.2%	RPMI	Mackay, L., Dodd, S., Dougall, I., Tomlinson, W., Lordan, J., Fisher, A. and Corris, P.: Isolation and Characterisation of Human Pulmonary Microvascular Endothelial Cells from Patients with Severe Emphysema., Respir Res 14, 23, 2013
人	Human	肺内皮 / Lung endothelial	Collagenase Type 2: 0.2%	PBS	Comhair, S., Xu, W., Mavrakis, L., Aldred, M., Asosingh, K. and Erzurum, S.: Human Primary Lung Endothelial Cells in Culture., Am J Respir Cell Mol Biol 46, 723-30, 2012
人	Human	肺 / Lung	Neutral Protease: 2 u/ml； Collagenase/Dispase: 0.1%； Deoxyribonuclease I: 0.01%	DMEM	Fujino, N., Kubo, H., Suzuki, T., Ota, C., Hegab, A., He, M., Suzuki, S., Suzuki, T., Yamada, M., Kondo, T., Kato, H. and Yamaya, M.: Isolation of Alveolar Epithelial Type II Progenitor Cells from Adult Human Lungs., Lab Invest 91, 363, 2011
人	Human	肺，成纤维细胞 / Lung，Fibroblasts	Trypsin: 0.05%	DMEM	Zhu, Y., Skold, C., Liu, X., Wang, H., Kohyama., T., Wen, F., Ertl, R., and Rennard, S.: Fibroblasts and Monocyte Macrophages Contract and Degrade Three-Dimensional Collagen Gels in Extended Co-Culture, Respir Res 2 (5), 295, 2001
人	Human, 16-24 week therapeutic abortions	肺泡II型，胎儿 / Alveolar type II，Fetal	Trypsin: 50 ug/ml	Ham's F-12, Eagle's MEM	Liley, H., Ertsey, R., Gonzales, L., Odom, M., Hawgood, S., Dobbs, L., and Ballard, P.: Synthesis of Surfactant Components by Cultured Type II Cells From Human Lung, Biochim Biophys Acta 961, 86, 1988
人	Human	上皮 / Epithelial	Trypsin: 0.02%	Medium 199	Lechner, J., Haugen, A., McClendon, I., and Pettis, E.: Clonal Growth of Normal Adult Human Bronchial Epithelial Cells in a Serum-Free Medium, In Vitro 18 (7), 633, 1982
人	Human, 12-16 wk old embryo	成纤维细胞 / Fibroblasts	Trypsin: 0.01%	Eagle's MEM	Kan, M., and Yamane, I.: In Vitro Proliferation and Lifespan of Human Diploid Fibroblasts in Serum-Free BSA-Containing Medium, J Cell Physiol 111, 155, 1982
人	Human fetuses, 80 day (also swine fetuses, 70 day, adult Amer Dutch, 250 day)	肺 / Lung	Collagenase: 0.01%	HBSS	Hinz, R., and Syverton, J.: Mammalian Cell Cultures for Study of Influenza Virus. I. Preparation of Monolayer Cultures with Collagenase, Proc Soc Exp Biol Med 101, 19, 1959
小鼠	Mouse	成纤维细胞 / Fibroblasts	Collagenase Type 2: 1.0% Deoxyribonuclease I: 0.001%	HBSS	Qiu, Y., Xu, Y., Wang, M., Miao, M., Zhou, H., Xu, J., Kong, J., Zheng, D., Li, R., Zhang, R., Guo, Y., Li, X., Cui, J., Qin, C. and Zhou , X.: Flavivirus Induces and Antagonizes Antiviral RNA Interference in Both Mammals and Mosquitoes., Sci. Adv. 6, 7989, 2020
小鼠	2-3 month	肺内皮 / Lung endothelial	Collagenase Type 1: 0.3%	DMEM	Wang, J., Niu, N., Xu, S. and Jin, Z.: A Simple Protocol for Isolating Mouse Lung Endothelial Cells., Sci Rep 9, 1458, 2019
小鼠	Mouse, 6 week				Alves CarlosRoberto, Souza RaquelSantos de, Charret KarenDos Santos, C?rtes LuziaMonteiro de Castro, Sá-Silva MatheusPereira de, Barral-Veloso Laura, Oliveira LuizFilipe Gon?alves, da Silva FranklinSouza: Understanding serine proteases implications on Leishmania spp lifecycle., Exp Parasitol 184, 67-81, 2018
小鼠	Mouse, 6 week	肺泡上皮 / Alveolar epithelial	Collagenase Type 1: 0.1%； Deoxyribonuclease I: 0.1%； Trypsin: 0.01%	DMEM/F-12	Lewis, K., Hall, J., Kiyotake, E., Christensen, T., Balasubramaniam, V. and Anseth, K.: Epithelial-Mesenchymal Crosstalk Influences Cellular Behavior in a 3D Alveolus-Fibroblast Model System., Biomaterials 155, 124, 2018
小鼠	Mouse	肺泡上皮 / Alveolar epithelial	Neutral Protease: 0.1%； Deoxyribonuclease I: 0.01%	DMEM	Sun, F., Xiao, G. and Qu, Z.: Isolation of Murine Alveolar Type II Epithelial Cells., Bio-protocol 7, , 2017
小鼠	Mouse	肺 / Lung	Collagenase Type 1: 0.15%； Deoxyribonuclease I: 0.04%	PBS	Yu, Y., O'Koren, E., Holten, D., Kan, M., Kojin, D., Nelson, E., Que, L. and Gunn, M.: A Protocol for the Comprehensive Flow Cytometric Analysis of Immune Cells in Normal and Inflammed Murine Non-Lymphoid Tissues, PLoS ONE 10, 1371, 2016
小鼠	Mouse	肺 / Lung	Collagenase Type 2: 0.3%； Deoxyribonuclease I: 0.3%	PBS	Braza, F., Dirou, S., Forest, V., Sauzeau, V., Hassoun, D., Chesne, J., Cheminant-Muller, M., Sagan, C., Magnan, A. and Lemarchand, P.: Mesenchymal Stem Cells Induce Suppressive Macrophages Through Phagocytosis in a Mouse Model of Asthma., Stem Cells 34, 1836-45, 2016
小鼠	Mouse, 8-12 week	上皮，内皮 / Epithelial, Endothelial	Collagenase Type 1: 0.5%； Deoxyribonuclease I: 0.1%； Neutral Protease: 0.1%	PBS	Singer, B., Mock, J., D'Alessio, F., Aggarwal, N., Mandke, P., Johnston, L. and Damarla, M.: Flow-Cytometric Method for Simultaneous Analysis of Mouse Lung Epithelial, Endothelial, and Hematopoietic Lineage Cells., Am J Physiol Lung Cell Mol Physiol 310, L796-801, 2016
小鼠	Mouse	肺泡 / Alveolar	Collagenase Type 1: 300 u/ml； Elastase: 4 u/ml； Neutral Protease: 5 u/ml； Deoxyribonuclease I: 200 u/ml	HBSS	Mohapatra, A., Van Dyken, S., Schneider, C., Nussbaum, J., Liang, H. and Locksley, R.: Group 2 Innate Lymphoid Cells Utilize the IRF4-IL-9 Module to Coordinate Epithelial Cell Maintenance of Lung Homeostasis., Mucosal Immunol , , 2015
小鼠	Mouse	肺 / Pulmonary	Collagenase Type 4: 0.16%； Deoxyribonuclease I: 0.1%	see reference	Kim, H., Lee, H., Chang, Y., Pichavant, M., Shore, S., Fitzgerald, K., Ikawura, Y., Israel, E., Bolger, K., Faul, J., DeKruyff, R. and Umetsu, D.: IL-17 Producing Innate Lymphoid Cells and the NLRP3 Inflammasone Facilitate Obescity-associated Airway Hyperractivity, Nat Med 20, 54, 2014
小鼠	Mouse, embryonic	腺泡祖细胞 / Avelolar progenitor	Elastase: 3 u/ml； Deoxyribonuclease I: 0.33 u/ml	DMEM/F12	Desai, T., Brownfield, D. and Krasnow, M.: Alveolar Progenitor and Stem Cells in Lung Development, Renewal and Cancer., Nature 507, 190-4, 2014
小鼠	Mouse	肺 / Lung	Collagenase Type 4: 0.3%	PBS	Guimond, D., Cam, N., Hirve, N., Duan, W., Lambris, J., Croft, M. and Tsoukas, C.: Regulation of Immune Responsiveness In Vivo by Disrupting an Early T-Cell Signaling Event Using a Cell-Permeable Peptide., PLoS ONE 8, e63645, 2013
小鼠	Mouse	肺 / Lung	Collagenase Type 4: 0.1%； Deoxyribonuclease I: 0.01%	HBSS	Misharin, A., Morales-Nebreda, L., Mutlu, G., Budinger, G. and Perlman, H.: Flow Cytometric Analysis of Macrophages and Dendritic Cell Subsets in the Mouse Lung., Am J Respir Cell Mol Biol 49, 503-10, 2013
小鼠	Mouse, 8-12 week	肺祖细胞 / Lung progenitor	Collagenase/Dispase: 0.2%； Neutral Protease: 5 u/ml； Deoxyribonuclease I: 0.0025%	DMEM	Driscoll, B., Kikuchi, A., Lau, A., Lee, J., Reddy, R., Jesudason, E., Kim, C. and Warburton, D.: Isolation and Characterization of Distal Lung Progenitor Cells., Methods Mol Biol 879, 109, 2012
小鼠	Mouse	肺肿瘤 / Lung tumor	Neutral Protease: 50 u/ml； Collagenase: 400 u/ml； Deoxyribonuclease I: 50 u/ml	DMEM	Vaughan, A., Halbert, C., Wootton, S. and Miller, A.: Lung Cancer in Mice Induced by the Jaagsiekte Sheep Retrovirus Envelope Protein is not Maintained by Rare Cancer Stem Cells, but Tumorigenicity does Correlate with Wnt Pathway Activation., Mol Cancer Res 10, 86, 2012
小鼠	Mouse	肺 / Lung	Collagenase Type 3: 0.1%； Deoxyribonuclease I: 0.0025%	RPMI 1640	Hardy, C., LeMasurier, J., Belz, G., Scalzo-Inguanti, K., Yao, J., Xiang, S., Kanellakis, P., Bobik, A., Strickland, D., Rolland, J., O'Hehir, R. and Plebanski, M.: Inert 50-nm Polystyrene Nanoparticles That Modify Pulmonary Dendritic Cell Function and Inhibit Allergic Airway Inflammation., J Immunol 188, 1431, 2012
小鼠	Mouse	肺引流 / Lung draining	Collagenase Type 4: 0.1-0.125%	HBSS	Rayamajhi, M., Redente, E., Condon, T., Gonzalez-Juarrero, M., Riches, D. and Lenz, L.: Non-Surgical Intratracheal Instillation of Mice with Analysis of Lungs and Lung Draining Lymph Nodes by Flow Cytometry., J Vis Exp 51, 2702, 2011
小鼠	Mouse	树突状 / Dendritic	Collagenase Type 1: 0.05%； Deoxyribonuclease I: 0.002%	HBSS	Lancelin, W. and Guerrero-Plata, A.: Isolation of Mouse Lung Dendritic Cells., J Vis Exp 57, e3563, 2011