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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

 

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
Bovine内皮,肺 / Endothelial,PulmonaryCollagenase: 1000 u/ml;
Collagenase Type 2: 1000 u/ml
PBS, CMFDel 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 IIElastase: 40 u/mlPBS (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 cellsTrypsin: 0.25%CF salt solutionPhillips, H.: Dissociation of Single Cells from Lung or Kidney Tissue with Elastase, In Vitro 8, 101, 1972
Human腺泡 / AveolarElastase: 12.9 u/mlPBSBahmed, 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
Human肺肿瘤 / Lung tumorCollagenase 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/F12Quatromoni, 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
Human肺泡上皮 / Alveolar epithelialNeutral Protease: 2 u/ml;
Trypsin: 0.25%;
Elastase: 10 u/ml
Bronchial Epithelial Growth MediumBarkauskas, 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
Human肺内皮 / Pulmonary endothelialCollagenase Type 2: 0.2%RPMIMackay, 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
Human肺内皮 / Lung endothelialCollagenase Type 2: 0.2%PBSComhair, 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
Human肺 / LungNeutral Protease: 2 u/ml;
Collagenase/Dispase: 0.1%;
Deoxyribonuclease I: 0.01%
DMEMFujino, 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
Human肺,成纤维细胞 / Lung,FibroblastsTrypsin: 0.05%DMEMZhu, 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
Human, 16-24 week therapeutic abortions肺泡II型,胎儿 / Alveolar type II,FetalTrypsin: 50 ug/mlHam's F-12, Eagle's MEMLiley, 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
Human上皮 / EpithelialTrypsin: 0.02%Medium 199Lechner, 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
Human, 12-16 wk old embryo成纤维细胞 / FibroblastsTrypsin: 0.01%Eagle's MEMKan, 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
Human fetuses, 80 day (also swine fetuses, 70 day, adult Amer Dutch, 250 day)肺 / LungCollagenase: 0.01%HBSSHinz, 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 / FibroblastsCollagenase Type 2: 1.0% Deoxyribonuclease I: 0.001%HBSSQiu, 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 endothelialCollagenase Type 1: 0.3%DMEMWang, 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 epithelialCollagenase Type 1: 0.1%;
Deoxyribonuclease I: 0.1%;
Trypsin: 0.01%
DMEM/F-12Lewis, 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 epithelialNeutral Protease: 0.1%;
Deoxyribonuclease I: 0.01%
DMEMSun, F., Xiao, G. and Qu, Z.: Isolation of Murine Alveolar Type II Epithelial Cells., Bio-protocol 7, , 2017
小鼠Mouse肺 / LungCollagenase Type 1: 0.15%;
Deoxyribonuclease I: 0.04%
PBSYu, 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 / LungCollagenase Type 2: 0.3%;
Deoxyribonuclease I: 0.3%
PBSBraza, 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, EndothelialCollagenase Type 1: 0.5%;
Deoxyribonuclease I: 0.1%;
Neutral Protease: 0.1%
PBSSinger, 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 / AlveolarCollagenase Type 1: 300 u/ml;
Elastase: 4 u/ml;
Neutral Protease: 5 u/ml;
Deoxyribonuclease I: 200 u/ml
HBSSMohapatra, 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 / PulmonaryCollagenase Type 4: 0.16%;
Deoxyribonuclease I: 0.1%
see referenceKim, 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 progenitorElastase: 3 u/ml;
Deoxyribonuclease I: 0.33 u/ml
DMEM/F12Desai, 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 / LungCollagenase Type 4: 0.3%PBSGuimond, 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 / LungCollagenase Type 4: 0.1%;
Deoxyribonuclease I: 0.01%
HBSSMisharin, 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 progenitorCollagenase/Dispase: 0.2%;
Neutral Protease: 5 u/ml;
Deoxyribonuclease I: 0.0025%
DMEMDriscoll, 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 tumorNeutral Protease: 50 u/ml;
Collagenase: 400 u/ml;
Deoxyribonuclease I: 50 u/ml
DMEMVaughan, 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 / LungCollagenase Type 3: 0.1%;
Deoxyribonuclease I: 0.0025%
RPMI 1640Hardy, 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 drainingCollagenase Type 4: 0.1-0.125%HBSSRayamajhi, 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 / DendriticCollagenase Type 1: 0.05%;
Deoxyribonuclease I: 0.002%
HBSSLancelin, W. and Guerrero-Plata, A.: Isolation of Mouse Lung Dendritic Cells., J Vis Exp 57, e3563, 2011


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