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：
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。
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| 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, Côrtes LuziaMonteiro de Castro, Sá-Silva MatheusPereira de, Barral-Veloso Laura, Oliveira LuizFilipe Gonçalves, 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, 8-10 week | 肺间充质干细胞 / Lung mesenchymal stem | Collagenase Type 2: 0.2% | HBSS | Chow, K., Jun, D., Helm, K., Wagner, D. and Majka, S.: Isolation & Characterization of Hoechst(low) CD45(negative) Mouse Lung Mesenchymal Stem Cells., J Vis Exp 56, e3159, 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 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase Type 1: 450 u/ml； Elastase: 4 u/ml； Neutral Protease: 5 u/ml； Deoxyribonuclease I: 0.33 u/ml |
DMEM/F12 | Rock, J, Barkauskas, C, Cronc, M, Xue, Y, Harris, J, Liang, J, Noble, P and Hogan, B.: Multiple Stromal Populations Contribute to Pulmonary Fibrosis Without Evidence for Epithelial to Mesenchymal Transition., Proc Natl Acad Sci U S A 108, E1475, 2011 |
| 小鼠 | Mouse, female, 6-12 week | 肺单核 / Lung mononuclear | Collagenase Type 4: 500 u/ml； Deoxyribonuclease I: 0.002% |
HBSS | Breslow, R., Rao, J., Xing, W., Hong, D., Barrett, N. and Katz, H.: Inhibition of Th2 Adaptive Immune Responses and Pulmonary Inflammation by Leukocyte Ig-Like Receptor B4 on Dendritic Cells., J Immunol 184, 1003, 2010 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase Type 2: 300 u/ml； Deoxyribonuclease I: 0.015% |
RPMI 1640 | Ferreira, C., Antunes, F., Leonard, V., Welstead, G., Richardson, C. and Cattaneo, R.: Measles Virus Infection of Alveolar Macrophages and Dendritic Cells Precedes Spread to Lymphatic Organs in Transgenic Mice Expressing Human Signaling Lymphocytic Activation Molecule (SLAM, CD150)., J Virol 84, 3033, 2010 |
| 小鼠 | Mouse, embryo, day 18 | 肺泡和成纤维细胞 / Alveolar and fibroblast | Trypsin: 2.5%； Deoxyribonuclease I: 0.2%； Collagenase Type 1: 1250 u/ml |
MEM | Trotter, A., Kipp, M., Schrader, R. and Beyer, C.: Combined Application of 17Beta-Estradiol and Progesterone Enhance Vascular Endothelial Growth Factor and Surfactant Protein Expression in Cultured Embryonic Lung Cells of Mice., Int J Pediatr 2009, 170491, 2009 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase Type 4: 0.1%； Deoxyribonuclease I: 0.01% |
HBSS | Zhao, J., Zhao, J., Van Rooijen, N. and Perlman, S.: Evasion by Stealth: Inefficient Immune Activation Underlies Poor T Cell Response and Severe Disease in SARS-CoV-Infected Mice., PLoS Pathog 5, e1000636, 2009 |
| 小鼠 | Mouse | 树突状 / Dendritic | Collagenase Type 1: 0.5% | HBSS | Flano, E., Jewell, N., Durbin, R. and Durbin, J.: Methods Used to Study Respiratory Virus Infection.,Curr Protoc Cell Biol Vol. Chapter 26,, , Unit 26.3, 2009 |
| 小鼠 | Mouse, 6-7 week | 白细胞和内皮细胞 / Leukocytes and endothelial | Collagenase: 0.2%； Deoxyribonuclease I: 40-80 u/ml |
PBS | Jungblut, M., Oeltze, K., Zehnter, I., Hasselmann, D. and Bosio, A.: Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue Using the GentleMACS Dissociator., J Vis Exp 29, , 2009 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase Type 2: 0.2% | PBS | Jones, R., Capen, D., Cohen, K., Munn, L., Jain, R. and Duda, D.: A Protocol for Phenotypic Detection and Characterization of Vascular Cells of Different Origins in a Lung Neovascularization Model in Rodents., Nat Protoc 3, 388-97, 2008 |
| 小鼠 | Mouse, 18-20 g | 肺 / Lung | Collagenase Type 3: 0.17% | HBSS | Ebeling, C., Lam, T., Gordon, J., Hollenberg, M. and Vliagoftis, H.: Proteinase-Activated Receptor-2 Promotes Allergic Sensitization to an Inhaled Antigen through a TNF-Mediated Pathway., J Immunol 179, 2910, 2007 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase Type 2: 300 u/ml； Deoxyribonuclease I: 0.001% |
Dulbecco's PBS | Finotto, S., Eigenbrod, T., Karwot, R., Boross, I., Doganci, A., Ito, H., Nishimoto, N., Yoshizaki, K., Kishimoto, T., Rose-John, S., Galle, P. and Neurath, M.: Local Blockade of IL-6R Signaling Induces Lung CD4+ T Cell Apoptosis in a Murine Model of Asthma Via Regulatory T Cells., Int Immunol 19, 685, 2007 |
| 小鼠 | Mouse, 6-16 week | 单核 / Mononuclear | Collagenase Type 4: 0.1% | RPMI 1640 | Abonia, J., Hallgren, J., Jones, T., Shi, T., Xu, Y., Koni, P., Flavell, R., Boyce, J., Austen, F. and Gurish, M.: Alpha-4 Integrins and VCAM-1, but not MAdCAM-1, are Essential for Recruitment of Mast Cell Progenitors to the Inflamed Lung., Blood 108, 1588-94, 2006 |
| 小鼠 | Mouse, 8-12 week | 单核细胞 / Mononuclear cells | Collagenase Type 1: 300 u/ml； Deoxyribonuclease I: 50 u/ml |
RPMI 1640 medium | Woolard MD, Hodge LM, Jones HP, Schoeb TR, and Simecka JW: The upper and lower respiratory tracts differ in their requirement of IFN-gamma and IL-4 in controlling respiratory mycoplasma infection and disease, J Immunol 172, 6875, 2004 |
| 小鼠 | Mouse, 6-10 week | 鼠肺内皮 / Murine pulmonary endothelial | Collagenase Type 2: 0.25% | HBSS | Andonegui G., Bonder C., Green F., Mullaly S., Zbytnuik L., Raharjo E., and Kubes P.: Endothelium-derived Toll-like Receptor-4 is the Key Molecule in LPS-induced Neutrophil Sequestration into Lungs, J Clin Invest 111, 1011, 2003 |
| 小鼠 | Mouse | 肺T淋巴细胞 / Pulmonary T lymphocytes | Collagenase Type 3: 10 mg/lung； Deoxyribonuclease I: 250 ug/lung |
RPMI medium | Huaux F, Liu T, McGarry B, Ullenbruch M, and Phan SH.: Dual roles of IL-4 in lung injury and fibrosis, J Immunol 170, 2083, 2003 |
| 小鼠 | Mouse | 肺泡上皮细胞 / Alveolar epithelial cells | Neutral Protease:； Deoxyribonuclease I: 0.01% |
DMEM | Paine R 3rd, Wilcoxen SE, Morris SB, Sartori C, Baleeiro CE, Matthay MA, and Christensen PJ: Transgenic overexpression of granulocyte macrophage-colony stimulating factor in the lung prevents hyperoxic lung injury, Am J Pathol 163, 2397, 2003 |
| 小鼠 | Mouse, 6-8 week | 肺和淋巴结细胞 / Lung and lymph node cells | Collagenase Type 2: 0.1%； Deoxyribonuclease I: 0.002% |
RPMI 1640 | Vermaelen KY, Carro-Muino I, Lambrecht BN, and Pauwels RA.: Specific migratory dendritic cells rapidly transport antigen from the airways to the thoracic lymph nodes, J Exp Med 193, 51, 2001 |
| 小鼠 | Mouse | 肺 / Lung | Collagenase: 100 u/ml； Deoxyribonuclease I: 200 u/ml |
Krebs-Henseleit Buffer | Freedman SD, Katz MH, Parker EM, Laposata M, Urman MY, and Alvarez JG.: A membrane lipid imbalance plays a role in the phenotypic expression of cystic fibrosis in cftr(-/-) mice, Proc Natl Acad Sci U S A 96, 13995, 1999 |
| 小鼠 | Mouse, 6-8 wk | 肺细胞 / Lung cells | Collagenase Type 3: 150 u/ml | HBSS | Stampfli MR, Wiley RE, Neigh GS, Gajewska BU, Lei XF, Snider DP, Xing Z, Jordana M: GM-CSF transgene expression in the airway allows aerosolized ovalbumin to induce allergic sensitization in mice, J Clin Invest 102, 1704-14, 1998 |
| 小鼠 | Mouse, female, 18-20 g | 鼠内皮细胞 / Murine endothelial cells | Collagenase Type 1: 1 mg/ml | DMEM | Dong QG, Bernasconi S, Lostaglio S, De Calmanovici RW, Martin-Padura I, Breviario F, Garlanda C, Ramponi S, Mantovani A,and Vecchi A.: A general strategy for isolation of endothelial cells from murine tissues. Characterization of two endothelial cell lines from the murine lung and subcutaneous sponge implants, Arterioscler Thromb Vasc Biol 17, 1599, 1997 |
| 小鼠 | Mouse, female | 抗原呈递细胞 / Antigen presenting cells | Collagenase: 150 u/ml； Deoxyribonuclease I: 30 u/ml |
RPMI 1640 | Hamilton-Easton A, and Eichelberger M: Virus-specific antigen presentation by different subsets of cells from lung and mediastinal lymph node tissues of influenza virus-infected mice, J Virol 69, 6359, 1995 |
| 猪 | Swine fetuses (70 day), adult Amer Dutch 250 day (also human fetuses, 80 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 |
| 兔 | Rabbit, New Zealand, white | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 0.05% | HBSS | Scott, J.: The Role of Sera, Growth Factors, and Hormones in the In Vitro Production of Disaturated Phosphatidylcholine and Propagation of Undifferentiated Type II Alveolar Cells from the Fetal Rabbit Lung, Exp Lung Res 12, 181, 1987 |
| 兔 | Rabbit, New Zealand white, adult,male | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 0.0025% | Joklik's MEM | Finkelstein, J., Maniscalco, W., and Shapiro, D.: Properties of Freshly Isolated Type II Alveolar Epithelial Cells, Biochim Biophys Acta 762, 398, 1983 |
| 兔 | Rabbit, New Zealand, male, 2-3 kg | 克拉拉细胞 / Clara cells | Protease: 0.1% | HEPES | Devereux, T., and Fouts, J.: Isolation and Identification of Clara Cells From Rabbit Lung, In Vitro 16 (11), 958, 1980 |
| 兔 | Rabbit, New Zealand, white, male, 1.7 Kg | 肺 / Lung | Pronase: 0.2% | Kreb's serum substitute solution, CMF | Gould, M., Clements, J., Jones, A., and Felts, J.: Dispersal of Rabbit Lung into Individual Viable Cells: A New Model for the Study of Lung Metabolism, Science 178, 1209, 1972 |
| 大鼠 | Rat | 肺 / Lung | Collagenase Type 2: 0.2% | PBS | Jones, R., Capen, D., Cohen, K., Munn, L., Jain, R. and Duda, D.: A Protocol for Phenotypic Detection and Characterization of Vascular Cells of Different Origins in a Lung Neovascularization Model in Rodents., Nat Protoc 3, 388-97, 2008 |
| 大鼠 | Rat, SD, 250-300g | 肺泡上皮 / Alveolar epithelial | Elastase: 3-4.5 u/ml | RPMI 1640 | Chen J., Chen Z., Narasaraju T., Jin N., and Liu L.: Isolation of Highly Pure Alveolar Epithelial Type I and Type II Cells from Rat Lungs, Lab Invest 84, 727, 2004 |
| 大鼠 | Rat, SD, 300-400 g | 肺内皮细胞 / Pulmonary endothelial cells | Collagenase Type 2: 1000 u/ml | DMEM/F-12 | King J, Hamil T, Creighton J, Wu S, Bhat P, McDonald F, and Stevens T: Structural and functional characteristics of lung macro- and microvascular endothelial cell phenotypes, Microvasc Res 67, 139, 2004 |
| 大鼠 | Rat, SD, 125-150 g | 肺泡类型 I 和 II / Alveolar type I & II | Elastase: 2.5-8 u/ml； Collagenase Type 1: 1.0% |
DMEM/F12 | Liebler, J., Borok, Z., Li, X., Zhou, B., Sandoval, A., Kim, K.and Crandall, E.: Alveolar Epithelial Type I Cells Express Beta 2-Adrenergic Receptors and G-protein Receptor Kinase 2., J Histochem Cytochem 52, 759, 2004 |
| 大鼠 | Rat, SD, 200-225 g | II型肺泡上皮细胞 / Type II alveolar epithelial cells | Elastase: 4.2 u/ml； Deoxyribonuclease I: 0.0001% |
DMEM | Sunil VR, Connor AJ, Guo Y, Laskin JD, and Laskin DL: Activation of type II alveolar epithelial cells during acute endotoxemia, Am J Physiol Lung Cell Mol Physiol 282, L872, 2002 |
| 大鼠 | Rat, SD, male, adult | 肺泡 / Alveolar | Elastase: 2.0 - 2.5 u/ml | DMEM/Ham's F-12 (see reference) | Kemp, P., Kim, K., Borok., Z., and Crandall, E.: Re-evaluating the Na+ Conductance of Adult Rat Alveolar Type II Pneumocytes: Evidence for the Involvement of cGMP-Activated Cation Channels, J Physiol 536 (3), 693, 2001 |
| 大鼠 | Rat, Lewis, male, 200-250g | 间质性肺巨噬细胞 / Interstitial lung macrophages | Collagenase Type 1: 100 u/ml； Deoxyribonuclease I: 50 u/ml |
RPMI-1640 | Steinmuller C, Franke-Ullmann G, Lohmann-Matthes ML, Emmendorffer A: Local activation of nonspecific defense against a respiratory model infection by application of interferon-gamma: comparison between rat alveolar and interstitial lung macrophages, Am J Respir Cell Mol Biol 22, 481-90, 2000 |
| 大鼠 | Rat, male | 肺泡上皮 / Alveolar epithelial | Elastase: 40 u/ml | DMEM | Planus, E., Galiacy, S., Matthay, M., Laurent, V., Gavrilovic, J., Murphy, G., Clerici, C., Isabey, D., Lafuma, C., and d'Ortho, M.: Role of Collagenase in Mediating in Vitro Alveolar Epithelial Wound Repair, J Cell Sci 112, 243, 1999 |
| 大鼠 | Rat, male, 200-250g | 肺动脉肌细胞 / Pulmonary arterial myocytes | Collagenase: 0.15%； Papain: 0.15%； Elastase: 0.05% |
PBS | Bakhramov, A, Evans, M and Kozlowski, R: Differential effects of hypoxia on the intracellular Ca2+ concentration of myocytes isolated from different regions of the rat pulmonary arterial tree, Exp Physiol 83, 337-47, 1998 |
| 大鼠 | Rat, embryonic day 15 | 胎儿肺泡上皮Ⅱ型 / Fetal alveolar epithelial type II | Trypsin: 0.1% | DMEM | Fraslon-Vanhulle, C, Bourbon, J and Batenburg, J: Culture of Fetal Alveolar Epithelial Type II Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 2,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 13A:2.1, 1995 |
| 大鼠 | Rat, SD, male, 125-175 g | 肺泡上皮 / Alveolar epithelial | Elastase: 2 u/ml | EBSS | Kim, K., Suh, D., Lubman, R., Danto, S., Borok, Z., and Crandall, E.: Studies on the Mechanisms of Active Ion Fluxes Across Alveolar Epithelial Cell Monolayers, J Tiss Cul Meth 14, 187, 1992 |
| 大鼠 | Rat, Wistar | 上皮 / Epithelial | Trypsin: 0.1% | HBSS | Jassal, D., Han, R., Caniggia, I., Post, M., and Tanswell, A.: Growth of Distal Fetal Rat Lung Epithelial Cells in a Defined Serum-Free Medium, In Vitro Cell Dev Biol 27A, 625, 1991 |
| 大鼠 | Rat, SD, 8 day | 间质 / Interstitial | Trypsin: 1.125% | HEPES buffer | Berk, J., Franzblau, C., and Goldstein, R.: Recombinant Interleukin-1beta Inhibits Elastin Formation by a Neonatal Rat Lung Fibroblast Subtype, J Biol Chem 266, 3192, 1991 |
| 大鼠 | Rat, Wistar, female, virgin | 肺泡上皮Ⅱ型 / Alveolar epithelial type II | Trypsin: 1% | Eagle's MEM | Fraslon, C., Rolland, G., Bourbon, J., Rieutort, M., and Valenza, C.: Culture of Fetal Alveolar Epithelial Type II Cells in Serum-Free Medium, In Vitro Cell Dev Biol 27A, 843, 1991 |
| 大鼠 | Rat, Fischer 344, male | 肺泡II型肺细胞 / Alveolar type II pneumocytes | Elastase: 40 u/ml | HEPES buffer (see reference) | Mangum, J., Everitt, J., Bonner, J., Moore, L., and Brody, A.: Co-Culture of Primary Pulmonary Cells to Model Alveolar Injury and Translocation of Proteins, In Vitro Cell Dev Biol 26, 1135, 1990 |
| 大鼠 | Rat, SD, male, 250-400 g | 肺泡Ⅱ型 / Alveolar type II | Elastase: 40 u/ml | Phosphate-buffered medium (see reference) | Ma, J., LaCagnin, L., Bowman, L., and Miles, P.: Carbon Tetrachloride Inhibits Synthesis of Pulmonary Surfactant Disaturated Phosphatidylcholines and ATP Production in Alveolar Type II Cells, Biochim Biophys Acta 1003, 136, 1989 |
| 大鼠 | Rat, Wistar, adult, male and pregnant female (known gestation) | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 0.1% | RPMI 1640 | Batenburg, J., Otto-Verberne, C., Have-Opbroek, A., and Klazinga, W.: Isolation of Alveolar Type II Cells from Fetal Rat Lung by Differential Adherence in Monolayer Culture, Biochim Biophys Acta 960, 441, 1988 |
| 大鼠 | Rat, SD, male, 150 - 200 g | II型肺细胞 / Pneumocytes type II | Trypsin: 0.30% | BSS | Brown, L. and Longmore, W.: Altered Phospholipid Secretion in Type II Pneumocytes Isolated from Streptozotocin-diabetic Rats, Biochim Biophys Acta 878, 258, 1986 |
| 大鼠 | Rat, SD, male, 180-200 g | 肺泡Ⅱ型 / Alveolar type II | Elastase: 4.3 u/ml | HEPES (see reference) | Dobbs, L., Gonzalez R., and Williams, M.: An Improved Method for Isolating Type ll Cells in High Yield and Purity, Am Rev Respir Dis 134, 141, 1986 |
| 大鼠 | Rat, SD, male | 肺泡Ⅰ型 / Alveolar type I | Trypsin: 0.05% | DMEM | Weller, N., and Karnovksy, M.: Isolation of Pulmonary Alveolar Type I Cells From Adult Rats, Am J Pathol 124, 448, 1986 |
| 大鼠 | Rat, fetus, 19 day | 成纤维细胞和 II 型 / Fibroblasts & type II | Trypsin: 0.05% | HBSS: serum free MEM, CMF | Post, M., Torday, J., and Smith, B.: Alveolar Type II Cells Isolated from Fetal Rat Lung Organotypic Cultures Synthesize and Secrete Surfactant-Associated Phospholipids and Respond to Fibroblast-Pneumonocyte Factor, Exp Lung Res 7, 53, 1984 |
| 大鼠 | Rat, SD, male, 150-400 g | 肺泡Ⅱ型 / Alveolar type II | Elastase: 4 u/ml | Auto-Pow Eagle's modified MEM | Goodman, B., Fleischer, R., and Crandall, E.: Evidence for Active Na+ Transport by Cultured Monolayers of Pulmonary Alveolar Epithelial Cells, Am J Physiol 245, C78, 1983 |
| 大鼠 | Rat, SD, male/female, 180-250 g | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 0.30% | BSS | Mason, R., Williams, M., Greenleaf, R., and Clements, J.: Isolation and Properties of Type II Alveolar Cells from Rat Lung, Am Rev Respir Dis 115, 1015, 1977 |
| 大鼠 | Rat, Wistar, pathogen free | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 0.50% | Earle's MEM | King, R.: Metabolic Fate of the Apoproteins of Pulmonary Surfactant, Am Rev Respir Dis 115, 73, 1977 |
| 大鼠 | Rat, fetus, 19 days | 肺泡肺细胞，II型 / Alveolar pneumonocytes, type II | Trypsin: 0.1 % | HBSS, CMF | Douglas, W., and Teel, R.: An Organotypic in Vitro Model System for Studying Pulmonary Surfactant Production by Type II Alveolar Pneumonocytes, Am Rev Respir Dis 113, 17, 1976 |
| 大鼠 | Rat, adult | 肺 / Lung | Collagenase: 0.1% | Moscona saline, CMF | Douglas, W., and Kaighn, M.: Clonal Isolation of Differentiated Rat Lung Cells, In Vitro 10, 230, 1974 |
| 大鼠 | Rat, SD, male, 100 g | 肺泡Ⅱ型 / Alveolar type II | Trypsin: 1.0% | Joklik's medium | Kikkawa, Y., and Yoneda, K.: Type II Epithelial Cell of the Lung. I. Method of Isolation, Lab Invest 30, 76, 1974 |
