原代细胞分离中，针对肿瘤涉及到肿瘤，羊膜间充质干细胞、肿瘤浸润淋巴细胞、上皮成釉细胞瘤，类器官，脑肿瘤和胰腺癌等等的解离通常会使用酶，如何选择合适的酶来完成对应肿瘤的解离，以及原代细胞采集的常规操作， 艾美捷针对肿瘤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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| 干细胞 | |||||
| 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 |
| 仓鼠 | Hamster, 90-100 g | 肿瘤 / Tumor | Hyaluronidase: 0.1% | Waymouth's MB | Gonzalez, A., Oberley, T., Schultz, J., Ostrom, J., and Li, J.: In Vitro Charactrization of Estrogen Induced Syrian Hamster Renal Tumors: Comparison with an Immortalized Cell Line Derived from Diethylstilbestrol-Treated Adult Hamster Kidney, In Vitro Cell Dev Biol 29A, 562, 1993 |
| 仓鼠 | Hamster, 6 week old | 颊袋 / Buccal pouch | Neutral Protease: 0.24% | CMF HBSS | Min, B., Kim, K., Cherrick, H., and Park, N.: Three Cell Lines from Hamster Buccal Pouch Tumors Induced by Topical 7,12-Dimethylbenz(a)Anthracene, Alone or in Conjunction with Herpes Simplex Virus Inoculation, In Vitro Cell Dev Biol 27A, 128, 1991 |
| 人 | Human | 肿瘤浸润淋巴细胞 / Tumor infiltrating lympocytes | Collagenase Type 1: 200 u/ml | RPMI | Jeske, S., Schuler, P., Doescher, J., Theodoraki, M., Laban, S., Brunner, C., Hoffman, T. and Wigand, M.: Age-related changes in T lymphocytes of patients with head and neck squamous cell carcinoma, Immun Ageing 17, 3, 2020 |
| 人 | Human | 肿瘤 / Tumor | Collagenase Type 4: 200 u/ml | DMEM | Phan, N., Hong, J., Tofig, B., Mapua, M., Elashoff, D., Moatamed, N., Huang, J., Memarzadeh, S., Damoiseaux, R. and Soragni, A.: A Simple High-Throughput Approach Identifies Actionable Drug Sensitivities in Patient-Derived Tumor Organoids., Commun Biol 2, 78, 2019 |
| 人 | Human | 成纤维细胞 / Fibroblasts | Collagenase Type 1: 0.5%； Neutral Protease: 0.1%； Deoxyribonuclease I: 1,000 u/ml |
FM Fibroblast media | Ayuso, J., Vitek, R., Swick, A., Skala, M., Wisinski, K., Kimple, R., Lambert, P. and Beebe, D.: Effects of Culture Method on Response to EGFR Therapy in Head and Neck Squamous Cell Carcinoma Cells, Sci Rep 9, 12480, 2019 |
| 人 | 类器官 / Organoids | Collagenase Type 4: 200-300 u/ml | DMEM | Neal, J,, Li ,X., Zhu, J., Giangarra, V., Grzeskowiak, C., Ju, J., Liu, I. Chiou, S., Salahudeen, A. and Smith, A. et al: Organoid Modeling of the Tumor Immune Microenvironment., Cell 175, 1972, 2018 | |
| 人 | Human | 上皮成釉细胞瘤 / Ameloblastoma epithelial | Collagenase: 0.1%； Neutral Protease: 50 u/ml |
MEM | Jiang, C., Zhang, Q., Shanti, R., Shi, S., Chang, T., Carrasco, L., Alawi, F. and Le, A.: Mesenchymal Stromal Cell-Derived Interleukin-6 Promotes Epithelial-Mesenchymal Transition and Acquisition of Epithelial Stem-Like Cell Properties in Ameloblastoma Epithelial Cells., Stem Cells 35, 2083-2094, 2017 |
| 人 | Human, 25-81 yr | 脑肿瘤 / Brain tumor | Neutral Protease: 0.11 u/ml； Collagenase Type 4: 0.05%； Hyaluronidase: 1,000 u/ml； Deoxyribonuclease I: 5 u/ml |
HBSS | Volovitz, I, Shapira, N., Ezer, H., Gafni, A., Lustgarten, M., Alter, T., Ben-Horin, I., Barzilai, O., Shahar, T., Kanner, A., Fried, I., Veshchev, I., Grossman, R. and Ram, Z,: A Non-Aggressive, Highly Efficient, Enzymatic Method for Dissociation of Human Brain-Tumors and Brain-Tissues to Viable Single-Cells., BMC Neurosci 17, 30, 2016 |
| 人 | Human | 胰腺癌 / Pancreatic cancer | Stemxyme 1: 0.2% | HBSS | Pham, K., Delitto, D., Knowlton, A., Hartlage, E., Madhavan, R., Gonzalo, D., Thomas, R., Behrns, K., George, T., Hughes, S., Wallet, S., Liu, C., Trevino, J.: Isolation of Pancreatic Cancer Cells from a Patient-Derived Xenograft Model Allows for Practical Expansion and Preserved Heterogeneity in Culture., Am J Pathol 186, , 2016 |
| 人 | Human | 非小细胞肺肿瘤 / Non-small cell lung tumor | Collagenase Type 1: 0.1%； Collagenase Type 2: 0.25% |
RPMI-1640 | Liang, S., Marti, T., Dorn, P., Froment, L., Hall, S., Berezowska, S., Kocher, G., Schmid, R. and Peng, R.: Blocking the Epithelial-to-Mesenchymal Transition Pathway Abrogates Resistance to Anti-Folate Chemotherapy in Lung Cancer., Cell Death Dis 6, e1824, 2015 |
| 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 | 肿瘤浸润淋巴细胞 / Tumour infiltrating lymphocytes | Collagenase: 1%； Hyaluronidase: 1%； Deoxyribonuclease I: 3,000 u/ml |
RPMI 1640 | Baldan, V., Griffiths, R., Hawkins, R. and Gilham, D: Efficient and Reproducible Generation of Tumour-Infiltrating Lymphocytes for Renal Cell Carcinoma, Br J Cancer 112, 1510, 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 | 胶质母细胞瘤 / Glioblastoma tumor | Trypsin: 0.025%； Collagenase Type 4: 500 u/ml |
HBSS | Hasselbach, L., Irtenkauf, S., Lemke, N., Nelson, K., Berezovsky, A., Carlton, E., Transou, A., Mikkelsen, T. and deCarvalho, A.: Optimization of High Grade Glioma Cell Culture from Surgical Specimens for Use in Clinically Relevant Animal Models and 3D Immunochemistry., J Vis Exp , e51088, 2014 |
| 人 | Human | 胶质瘤癌干细胞 / Glioma cancer stem | PDS kit: per instructions | Neurobasal | Yan, K., Wu, Q., Yan, D., Lee, C., Rahim, N., Tritschler, I., DeVecchio, J., Kalady, M., Hjelmeland, A. and Rich, J.: Glioma Cancer Stem Cells Secrete Gremlin1 to Promote their Maintenance Within the Tumor Hierarchy., Genes Dev 28, 1085-100, 2014 |
| 人 | Human | 黑色素瘤 / Melanoma | Collagenase Type 4: 0.1%； Deoxyribonuclease I: 0.01% |
PBS | Welte, Y., Davies, C., Schafer, R. and Regenbrecht, C.: Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma., J Vis Exp 73, e50200, 2013 |
| 人 | Human | 肿瘤 / Tumor | Collagenase Type 1: 0.1%； Hyaluronidase: 0.01%； Deoxyribonuclease I: 0.01% |
DMEM/F12 | Chou, J., Fitzgibbon, M., Mortales, C., Towlerton, A., Upton, M., Yeung, R., McIntosh, M. and Warren, E.: Phenotypic and Transcriptional Fidelity of Patient-Derived Colon Cancer Xenografts in Immune-Deficient Mice., PLoS ONE 8, e79874, 2013 |
| 人 | Human | 癌干细胞 / Cancer stem | Trypsin: 0.25%； Collagenase: 0.05%； Hyaluronidase: 0.005% |
RPMI | Dobbin, Z. and Landen, C.: Isolation and Characterization of Potential Cancer Stem Cells from Solid Human Tumors--Potential Applications., Curr Protoc Pharmacol 63, Unit 14.28., 2013 |
| 人 | Human | 大肠癌 / Colorectal cancer | Collagenase Type 4: 2%； Deoxyribonuclease I: 0.1% |
RPMI 1640 | Zhou, J, Belov, L., Solomon, M., Chan, C., Clarke, S. and Christopherson, R.: Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing., J Vis Exp 55, e3322, 2011 |
| 人 | Human | 脑肿瘤 / Brain tumor | Collagenase Type 1: 0.04%；
Hyaluronidase: 0.01%； Deoxyribonuclease I: 0.02%； Neutral Protease: 0.008%； |
DMEM/F12 | Hussein, D., Punjaruk, W., Storer, L., Shaw, L., Othman, R., Ottoman, R., Peet, A., Miller, S., Bandopadhyay, G., Heath, R., Kumari, R., Bowman, K., Braker, P., Rahman, R., Jones, G., Watson, S. and Lowe, J.: Pediatric Brain Tumor Cancer Stem Cells: Cell Cycle Dynamics, DNA Repair, and Etoposide Extrusion., Neuro Oncol 13, 70-83, 2011 |
| 人 | Human | 致瘤性黑色素瘤 / Tumorigenic melanoma | Collagenase Type 4: 200 u/ml； Trypsin: 0.05%； Deoxyribonuclease I: 50-100 u/ml |
PBS | Quintana, E., Shackleton, M., Foster, H., Fullen, D., Sabel, M., Johnson, T. and Morrison, S.: Phenotypic Heterogeneity Among Tumorigenic Melanoma Cells from Patients that is Reversible and Not Hierarchically Organized., Cancer Cell 18, 510, 2010 |
| 人 | Human | 胰腺肿瘤 / Pancreatic tumor | Collagenase Type 4: 200 u/ml | RPMI-1640 | Kim, M., Evans, D., Wang, H., Abbruzzese, J., Fleming, J. and Gallick, G.: Generation of Orthotopic and Heterotopic Human Pancreatic Cancer Xenografts in Immunodeficient Mice., Nat Protoc 4, 1670, 2009 |
| 人 | Human | 肿瘤 / Tumor | Collagenase Type 4: 0.1%； Hyaluronidase: 0.07%； Deoxyribonuclease I: 0.04% |
see reference | Sauvageot, C., Weatherbee, J., Kesari, S., Winters, S., Barnes, J., Dellagatta, J., Ramakrishna, N., Stiles, C., Kung, A., Kieran, M. and Wen, P.: Efficacy of the HSP90 Inhibitor 17-AAG in Human Glioma Cell Lines and Tumorigenic Glioma Stem Cells.,Neuro Oncol Vol. 11,, , 109, 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 | 乳腺上皮 / Breast epithelial | Collagenase Type 3: 200 u/ml | HBSS | Liu, R., Wang, X., Chen, G., Dalerba, P., Gurney, A., Hoey, T., Sherlock, G., Lewicki, J., Shedden, K. and Clarke, M.: The Prognostic Role of a Gene Signature from Tumorigenic Breast-Cancer Cells., N Engl J Med 356, 217, 2007 |
| 人 | Human | 前列腺基质细胞 / Prostate stromal cells | Collagenase Type 1: 0.1% | RPMI 1640 | Nakashiro Koh-Ichi, Hara Shingo, Shinohara Yuji, Oyasu Miho, Kawamata Hitoshi, Shintani Satoru, Hamakawa Hiroyuki, Oyasu Ryoichi: Phenotypic switch from paracrine to autocrine role of hepatocyte growth factor in an androgen-independent human prostatic carcinoma cell line, CWR22R, Am J Pathol 165, 533-40, 2004 |
| 人 | Human, adult | 人类滑膜肉瘤 / Human synovial sarcoma | Collagenase Type 2: 200 u/ml | DMEM/F-12 | Nishio Jun, Iwasaki Hiroshi, Ishiguro Masko, Ohjimi Yuko, Fujita Chikako, Isayama Teruto, Naito Masatoshi, Oda Yoshinao, Kaneko Yasuhiko, Kikuchi Masahiro: Establishment of a new human synovial sarcoma cell line, FU-SY-1, that expresses c-Met receptor and its ligand hepatocyte growth factor, Int J Oncol 21, 17-23, 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 | 上皮细胞，成纤维细胞 / Epithelial, fibroblasts | Trypsin: 0.25% | Ham's F-12 | MacLeod, R: Rapid Monolayer Primary Cell Culture from Tissue Biopsy,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons Ltd, 3E:2.1, 1995 |
| 人 | Human | 结肠腺癌 / Colon adenocarcinoma | Hyaluronidase: 100 u/ml | DMEM | Hague, A and Paraskeva, C: Colon Adenocarcinoma Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12C:1.1, 1995 |
| 人 | Human | 肿瘤，乳房 / Tumor, breast | Hyaluronidase: 100 u/ml | DMEM | Beaupain, R., Mainquene, C., Brouty-Boye, D., Planchon, P., and Magniew, V.: "Normal" Breast Cells Adjacent to a Tumor Grown in Long-term Three Dimensional Culture, In Vitro Cell Dev Biol 29, 100, 1993 |
| 人 | Human | 胶质瘤 / Glioma | Hyaluronidase: 0.01% | HBSS | Kruse, C., Mitchell, D., Kleinschmidt-DeMasteis, B., Franklin, W., Morse, H., Spector, E., and Lillehei, K.: Characterization of a Continuous Human Glioma Cell Line DBTRG-OSMG: Growth Kinetics, Karyotype, Receptor Expression and Tumor Suppressor Gene Analyses, In Vitro Cell Dev Biol 28, 609, 1992 |
| 人 | Human | 上皮 / Epithelial | Collagenase: 2.0% | DMEM/Ham's F-12 | Emerman, J. and Wilkinson, D.: Routine Culturing of Normal, Dysplastic and Malignant Human Mammary Epithelial Cells from Small Tissue Samples, In Vitro Cell Dev Biol 26, 1186, 1990 |
| 人 | Human | 肿瘤 / Tumor | Neutral Protease: 0.24% | DMEM/Ham's F-12 | Boyd, J., Rinehart Jr., C., Walton, L., Siegal, G. and Kaufman, D.: Ultrastructural Characterization of Two New Human Endometrial Carcinoma Cell Lines and Normal Human Endometrial Epithelial Cells Cultured on Extracellular Matrix, In Vitro Cell Dev Biol 26, 701, 1990 |
| 人 | Human, 9-74 year | 神经纤维瘤 / Neurofibroma | Neutral Protease: 1.25 u/ml； Collagenase Type 1: 0.05%； Hyaluronidase: 0.1% |
L-15 | Sheela S, Riccardi VM, Ratner N: Angiogenic and invasive properties of neurofibroma Schwann cells, J Cell Biol 111, 645-53, 1990 |
| 人 | Human | 肿瘤 / Tumor | Trypsin: 0.05% | DMEM | Sacks, P., Parnes, S., Gallick, G., Mansouri, Z., Lichtner, R., Satya-Prakash, K., Pathak, S, and Parsons, D.: Establishment and Characterization of Two New Squamous Cell Carcinoma Cell Lines Derived from Tumors of the Head and Neck, Cancer Res 48, 2858, 1988 |
| 人 | Human | 肿瘤，结肠 / Tumor, colon | Trypsin: 0.25% | McCoy's | Brattain, M., Marks, M., McCombs, J., Finely, W., and Brattain, D.: Characterization of Human Colon Carcinoma Cell Lines Isolated From a Single Primary Tumour, Br J Cancer 47, 373, 1983 |
| 人 | Human | 上皮和肿瘤，结肠 / Epthelial and tumor，Colon | Collagenase: 300 u/ml | PBS medium 199 or medium F 12 | Friedman, E., Higgins, P., Lipkin, M., Shinya, H., and Gelb, A.: Tissue Culture of Human Epithelial Cells from Benign Colonic Tumors, In Vitro 17, 632, 1981 |
| 人 | Human | 肿瘤，乳房 / Tumor, breast | Neuraminidase: 0.8 u/ml | HBSS | Leung, C., and Shiu, R.: Morphological and Proliferative Characteristics of Human Breast Tumor Cells Cultured on Plastic and in Collagen Matrix, In Vitro 18, 476, 1981 |
| 人 | Human | 黑色素瘤，转移性肿瘤 / Melanoma，Metastatic tumors | Collagenase Type 3: 0.10% | DMEM | Creasey, A., Smith, H., Hackett, A., Fukuyama, K., Epstein, W., and Madin, S.: Biological Properties of Human Melanoma Cells in Culture, In Vitro 15, 342, 1979 |
| 人 | Human | 乳腺肿瘤，硬 / Mammary tumors, hard | Collagenase: 0.10% | RPMI-1640 w/ 5% Fetal Calf Serum | Lasfargues, E.: Human Mammary Tumors,Tissue Culture Methods/Applications,Kruse, P., and Patterson, M., Academic Press, 45, 1973 |
| 小鼠 | Mouse | 肝肿瘤 / liver tumor | Collagenase Type 2: 0.25% | HBSS | Walker, S., Wankell, M., Ho, V., White, R., Deo, N., Devine, C., Dewdney, B., Bhathal, P., Govaere, O., Roskams, T., Qiao, L., George, J. and Hebbard, L.: Targeting mTOR and Src Restricts Hepatocellular Carcinoma Growth in a Novel Murine Liver Cancer Model., PLoS ONE 14, e0212860, 2019 |
| 小鼠 | Mouse, adult | 类器官 / Organoids | Collagenase Type 4: 200-300 u/ml | DMEM | Neal, J,, Li ,X., Zhu, J., Giangarra, V., Grzeskowiak, C., Ju, J., Liu, I. Chiou, S., Salahudeen, A. and Smith, A. et al: Organoid Modeling of the Tumor Immune Microenvironment., Cell 175, 1972, 2018 |
| 小鼠 | Mouse | 肿瘤来源的中性粒细胞瘤 / Tumor derived neutroblastoma | Papain: 10 u/ml； Deoxyribonuclease I: 250 u/ml |
DPBS | Erdreich-Epstein, A., Singh, A., Joshi, S., Vega, F., Guo, P., Xu, J., Groshen, S., Ye, W., Millard, M., Campan, M., Morales, G., Garlich, J., Laird, P., Seeger, R., Shimada, H. and Durden, D.: Association of High Microvessel A,B and Low PTEN with Poor Outcome in Stage 3 Neuroblastoma: Rationale for Using First in Class Dual PI3K/BRD4 Inhibitor, SF1126., Oncotarget 8, 52193-52210, 2017 |
| 小鼠 | Mouse | 胶质母细胞瘤 / Glioblastoma | PDS kit: per instructions | DMEM/F12 | Boyd, N., Walker, K., Fried, J., Hackney, J., McDonald, P., Benavides, G., Spina, R., Audia, A., Scott, S., Libby, C., Tran, A., Bevensee, M., Griguer, C., Nozell, S., Gillespie, G. and Nabors, B.: Addition of Carbonic Anhydrase 9 Inhibitor SLC-0111 to Temozolomide Treatment Delays Glioblastoma Growth In Vivo., JCI Insight 2, , 2017 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase Type 4: 0.1%； Neutral Protease: 0.1%； Deoxyribonuclease I: 0.02% |
RPMI-1640 | Filtjens, J., Keirsse, J., Van Ammel, E., Taveirne, S., Van Acker, A., Kerre, T., Taghon, T., Vandekerckhove, B., Plum, J., Van Ginderachter, J. and Leclercq, G.: Expression of the Inhibitory Ly49E Receptor is Not Critically Involved in the Immune Response Against Cutaneous, Pulmonary or Liver tumours., Sci Rep 6, 30564, 2016 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase: 0.1%； Neutral Protease: 0.5 u/ml |
F12K | Aguilera, T., Rafat, M., Castellini, L., Shehade, H., Kariolis, M., Hui, A., Stehr, H., von Eyben, R., Jiang, D., Ellies, L., Koong, A., Diehn, M., Rankin, E. Graves, E. and Giaccia, A.: Reprogramming the Immunological Microenvironment through Radiation and Targeting Axl., Nat Commun 7, 13898, 2016 |
| 小鼠 | Mouse | 肿瘤浸润淋巴细胞 / Tumor-infiltrating lymphocytes | Collagenase Type 4: 0.25% | DMEM | Singer, M., Wang, C., Cong, L., Marjanovic, N., Kowalczyk, Mo., Zhang, H., Nyman, J., Sakuishi, K., Kurtulus, S., Gennert, D., Xia, J., Kwon, J., Nevin, J., Herbst, R., Yanai, I., Rozenblatt-Rosen, O. and Kuchroo, V.: A Distinct Gene Module for Dysfunction Uncoupled from Activation in Tumor-Infiltrating T Cells., Cell 166, 1500-1511.e9, 2016 |
| 小鼠 | Mouse | 胶质母细胞瘤 / Glioblastoma | PDS kit: per instructions | Neurobasal | Otvos, B., Silver, D., Mulkearns-Hubert, E., Alvarado, A., Turaga, S., Sorensen, M., Rayman, P., Flavahan, W., Hale, J., Stoltz, K., Sinyuk, M., Wu, Q., Jarrar, A., Kim, S., Fox, P., Nakano, I. and Rich, J.: Cancer Stem Cell-Secreted Macrophage Migration Inhibitory Factor Stimulates Myeloid Derived Suppressor Cell Function and Facilitates Glioblastoma Immune Evasion., Stem Cells 34, 2026-39, 2016 |
| 小鼠 | Mouse | 角质形成细胞 / Keratinocytes | Trypsin: 0.25%；
Collagenase Type 1: 0.12%； Collagenase Type 2: 0.05%； Collagenase Type 4: 0.05%； Hyaluronidase: 0.1%； |
Low calcium containing media | Blanco, S., Bandiera, R., Popis, M., Hussain, S., Lombard, P., Aleksic, J., Sajini, A., Tanna, H., Cortes-Garrido, R., Gkatza, N., Dietmann, S. and Frye, M.: Stem Cell Function and Stress Response are Controlled by Protein Synthesis., Nature 534, 335-40, 2016 |
| 小鼠 | Mouse | 乳腺肿瘤 / Mammary tumor | Collagenase Type 1: 200 u/ml； Hyaluronidase: 0.001% |
DMEM/F12 | Hosseini, H., Obradovic, M., Hoffmann, M., Harper, K., Sosa, M., Werner-Klein, M., Nanduri, L., Werno, C., Ehrl, C., Maneck, M., Patwary, N., Haunschild, G., Guzvic, M., Reimelt, C. et al.: Early Dissemination Seeds Metastasis in Breast Cancer., Nature , , 2016 |
| 小鼠 | Mouse | 内皮 / Endothelial | Collagenase Type 2: 0.1%； Neutral Protease: 0.25 u/ml； Deoxyribonuclease I: 0.0075% |
DMEM | Xiao, L., McCann, J. and Dudley, A.: Isolation and Culture Expansion of Tumor-specific Endothelial Cells., J Vis Exp , e53072, 2015 |
| 小鼠 | Mouse | 甲状腺肿瘤 / Thyroid tumor | Collagenase Type 1: 300 u/ml； Hyaluronidase: 100 u/ml； Deoxyribonuclease I: 0.01% |
DMEM | Kitajima, S., Kohno, S., Kondoh, A., Sasaki, N., Nishimoto, Y., Li, F., Mohammed, M., Muranaka, H., Nagatani, N., Suzuki, M., Kido, Y. and Takahashi, C.: Undifferentiated State Induced by Rb-p53 Double Inactivation in Mouse Thyroid Neuroendocrine Cells and Embryonic Fibroblasts., Stem Cells 33, 1657-69, 2015 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase Type 4: 0.1； Hyaluronidase: 0.01%； Deoxyribonuclease I: 30 u/ml |
PBS | Devaud, C., Westwood, J., John, L., Flynn, J., Paquet-Fifield, S., Duong, C., Yong, C., Pegram, H., Stacker, S., Achen, M. Stewart, T., Snyder, L., Teng, M., Smyth, M., Darcy, P. and Kershaw, M: Tissues in Different Anatomical Sites can Sculpt and Vary the Tumor Microenvironment to Affect Responses to Therapy., Mol Ther 22, 18-27, 2014 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase Type 1: 250 u/ml | DMEM | Ali, O., Verbeke, C., Johnson, C., Sands, R., Lewin, S., White, D., Doherty, E., Dranoff, G. and Mooney, D.: Identification of Immune Factors Regulating Antitumor Immunity Using Polymeric Vaccines with Multiple Adjuvants., Cancer Res 74, 1670-81, 2014 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase Type 2: 0.2%； Neutral Protease: 2.5 u/ml； Deoxyribonuclease I: 0.0075% |
DMEM | Dunleavey, J., Xiao, L., Thompson, J., Kim, M., Shields, J., Shelton, S., Irvin, D., Brings, V., Ollila, D., Brekken, R., Dayton, P., Melero-Martin, J. and Dudley, A.: Vascular Channels Formed by Subpopulations of PECAM1+ Melanoma Cells., Nat Commun 5, 5200, 2014 |
| 小鼠 | Mouse | 成纤维细胞 / Fibroblasts | Collagenase Type 2: 0.25%； Collagenase Type 4: 0.25%； Deoxyribonuclease I: 0.05% |
DMEM | Sharon, Y., Alon, L., Glanz, S., Servais, C. and Erez, N.: Isolation of Normal and Cancer-Associated Fibroblasts from Fresh Tissues by Fluorescence Activated Cell Sorting (FACS)., J Vis Exp 71, e4425, 2013 |
| 小鼠 | Mouse | 前列腺肿瘤 / Prostate tumor | Collagenase Type 4: 1,600 u/ml | DMEM/F12 | Mazzoleni, S., Jachetti, E., Morosini, S., Grioni, M., Piras, I., Pala, M., Bulfone, A., Freschi, M., Bellone, M. and Galli, R.: Gene Signatures Distinguish Stage-Specific Prostate Cancer Stem Cells Isolated From Transgenic Adenocarcinoma of the Mouse Prostate Lesions and Predict the Malignancy of Human Tumors., Stem Cells Transl Med 2, 678, 2013 |
| 小鼠 | Mouse | 乳腺肿瘤 / Mammary tumor | Collagenase: 0.15%； Hyaluronidase: 0.020% |
DMEM/F12 | Liu, X., Johnson, S., Liu, S., Kanojia, D., Yue, W., Singh, U., Wang, Q, Wang Qi, Nie, Q. and Chen H.: Nonlinear Growth Kinetics of Breast Cancer Stem Cells: Implications for Cancer Stem Cell Targeted Therapy., Sci Rep 3, 2473, 2013 |
| 小鼠 | Mouse | 横纹肉瘤肿瘤 / Rhabdomycosarcoma tumor | Trypsin: 0.012%； Collagenase Type 2: 0.1% |
DMEM | Chen, X., Stewart, E., Shelat, A., Qu, C., Bahrami, A., Hatley, M., Wu, G., Bradley, C., McEvoy, J., Pappo, A., Spunt, S., Valentine, M., Valentine, V., Krafcik, F., Lang, W., Wierdl, M. and Tsurkan, L.: Targeting Oxidative Stress in Embryonal Rhabdomyosarcoma., Cancer Cell 24, 710-24, 2013 |
| 小鼠 | Mouse | 肿瘤内皮 / Tumor endothelial | Collagenase Type 1: 0.2% | HBSS | Kazerounian, S., Gerald, D., Huang, M., Chin, R, Udayakumar, D, Zheng, N., O'Donnell, R., Perruzzi, C., Mangiante, L., Pourat, J., Phung, T., Bravo-Nuevo, A., Shechter, S., McNamara, S. and Duhadaway, K.: RhoB Differentially Controls Akt Function in Tumor Cells and Stromal Endothelial Cells During Breast Tumorigenesis., Cancer Res 73, 50, 2013 |
| 小鼠 | Mouse, male | 树突状，巨噬细胞 / Dendritic, macrophages | Collagenase (1 or 4): 100-200 u/ml； Deoxyribonuclease I: 0.01% |
RPMI | Watkins, S., Zhu, Z., Watkins, K. and Hurwitz, A.: Isolation of Immune Cells from Primary Tumors., J Vis Exp 64, e3952, 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 | 巨噬细胞 / Macrophages | Collagenase Type 4: 0.2% | PBS | Hamzah, J., Kotamraju, V., Seo, J., Agemy, L., Fogal, V., Mahakian, L., Peters, D., Roth, L., Gagnon, M., Ferrara, K. and Ruoslahti, E.: Specific Penetration and Accumulation of a Homing Peptide Within Atherosclerotic Plaques of Apolipoprotein E-deficient Mice., Proc Natl Acad Sci U S A 108, 7154, 2011 |
| 小鼠 | Mouse | 胰腺癌干细胞 / Pancreatic cancer stem | Collagenase Type 4: 200 u/ml； Neutral Protease: 0.6 u/ml |
DMEM | Rasheed, Z., Wang, Q. and Matsui, W.: Isolation of Stem Cells from Human Pancreatic Cancer Xenografts., J Vis Exp 43, 2169, 2010 |
| 小鼠 | Mouse | 肿瘤浸润淋巴细胞 / Tumor infiltration lymphocytes | Collagenase Type 1: 0.25%； Collagenase Type 2: 0.15%； Collagenase Type 4: 0.1%； Hyaluronidase: 0.025% |
RPMI 1640 | Kwong, B., Roberts, S., Silberzahn, T., Filler, R., Neustadter, J., Galan, A., Reddy, S., Lin, W., Ellis, P., Langford, C., Hayday, A. and Girardi, M.: Molecular Analysis of Tumor-Promoting CD8+ T cells in Two-Stage Cutaneous Chemical Carcinogenesis., J Invest Dermatol 130, 1726, 2010 |
| 小鼠 | Mouse, 4-6 week | 胰腺肿瘤 / Pancreatic tumor | Collagenase Type 4: 200 u/ml | RPMI-1640 | Kim, M., Evans, D., Wang, H., Abbruzzese, J., Fleming, J. and Gallick, G.: Generation of Orthotopic and Heterotopic Human Pancreatic Cancer Xenografts in Immunodeficient Mice., Nat Protoc 4, 1670, 2009 |
| 小鼠 | Mouse | 肿瘤 / Tumor | Collagenase Type 3: 200 u/ml | RPMI-1640 | Prince, M., Sivanandan, R., Kaczorowski, A., Wolf, G., Kaplan, M., Dalerba, P., Weissman, I., Clarke, M. and Ailles, L.: Identification of a Subpopulation of Cells with Cancer Stem Cell Properties in Head and Neck Squamous Cell Carcinoma., Proc Natl Acad Sci U S A 104, 973, 2007 |
| 小鼠 | Mouse, 30 week | 肿瘤内皮 / Tumor endothelial | Collagenase Type 4: 500 u/ml； Collagenase Type 2: 550 u/ml； Deoxyribonuclease I: 3 u/ml |
PBS | Berger, M., Bergers, G., Arnold, B., Hammerling, G. and Ganss, R.: Regulator of G-protein Signaling-5 Induction in Pericytes Coincides With Active Vessel Remodeling During Neovascularization, Blood 105, 1094, 2005 |
| 小鼠 | Mouse | 颗粒细胞前体，前肿瘤细胞和肿瘤细胞 / Granule cell precursors, pre-neoplastic and tumor cells | Papain: 10 u/ml； Deoxyribonuclease I: 250 u/ml |
Neurobasal/B27 | Oliver, T., Read, T., Kessler, J., Mehmeti, A., Wells, J., Huynh, T., Lin, S. and Wechsler-Reya, R.: Loss of Patched and Disruption of Granule Cell Development in a Pre-Neoplastic Stage of Medulloblastoma., Development 132, 2425, 2005 |
| 小鼠 | Mouse, 8-10 week | 肿瘤相关内皮细胞 / Tumor associated endothelial cells | Collagenase Type 2: | see reference | Hida Kyoko, Hida Yasuhiro, Amin Dhara N, Flint Alan F, Panigrahy Dipak, Morton Cynthia C, Klagsbrun Michael: Tumor-associated endothelial cells with cytogenetic abnormalities, Cancer Res 64, 8249-55, 2004 |
| 小鼠 | Mouse | 胰腺肿瘤 / Pancreatic tumor | Collagenase Type 2: 0.5%； Collagenase Type 4: 0.5%； Deoxyribonuclease I: 0.2% |
PBS | Bergers Gabriele, Song Steven, Meyer-Morse Nicole, Bergsland Emily, Hanahan Douglas: Benefits of targeting both pericytes and endothelial cells in the tumor vasculature with kinase inhibitors, J Clin Invest 111, 1287-95, 2003 |
| 小鼠 | Mouse | 肿瘤浸润淋巴细胞 / Tumor-infiltrating lymphocyte | Collagenase: 200 u/ml | RPMI 1640 | Uekusa Yasuhiro, Yu Wen-Gong, Mukai Takao, Gao Ping, Yamaguchi Nobuya, Murai Masako, Matsushima Kouji, Obika Satoshi, Imanishi Takeshi, Higashibata Yuji, Nomura Shintaro, Kitamura Yukihiko, Fujiwara Hiromi, Hamaoka Toshiyuki: A pivotal role for CC chemokine receptor 5 in T-cell migration to tumor sites induced by interleukin 12 treatment in tumor-bearing mice, Cancer Res 62, 3751-8, 2002 |
| 小鼠 | Mouse, male | 黑色素瘤细胞 / Melanoma tumor cells | Collagenase Type 2: 0.5% | DMEM | Arbiser JL, Raab G, Rohan RM, Paul S, Hirschi K, Flynn E, Price ER, Fisher DE, Cohen C, Klagsbrun M: Isolation of mouse stromal cells associated with a human tumor using differential diphtheria toxin sensitivity, Am J Pathol 155, 723-9, 1999 |
| 小鼠 | Mouse (BALB/cfC3H) | 乳腺肿瘤，上皮 / Mammary tumors，Epithelial | Collagenase: 1.0% | HBSS | Yang, J., Guzman, R., Richards, J., and Nandi, S.: Primary Cultures of Mouse Mammary Tumor Epithelial Cells Embedded in Collagen Gels, In Vitro 16, 502, 1980 |
| 小鼠 | Mouse, BALB/cfC3H/Crgl | 肿瘤，上皮肿瘤 / Neoplastic，Epithelial tumor | Trypsin: | DMEM | Hosick, H.: A Note on Growth Patterns of Epithelial Tumor Cells in Primary Culture, Cancer Res 34, 259, 1974 |
| 小鼠 | Mouse, lactating, 14 day | 乳腺 / Mammary | Trypsin NF 1:250: 0.25% | HBSS | Kopelovich, L., Abraham, S., McGrath, H., DeOme, K., Chaikoff, I.: Metabolic Characteristics of a Naturally Ocurring Preneoplastic Tissue. I. Glycolytic Enzyme Activators of Hyperplastic Alveolar Nodule Outgrowths and Adenocarcinomas of Mouse Mammary Gland, Cancer Res 26, 1534, 1966 |
| 大鼠 | Rat, adult | 肿瘤 / Tumor | Collagenase Type 1: 0.16%； Hyaluronidase: 0.002%； Deoxyribonuclease I: 0.006% |
DMEM/F12 | Duarte, S., Momier, D., Baque, P., Casanova, V., Loubat, A., Samson, M., Guigonis, J., Staccini, P., Saint-Paul, M., De Lima, M., Carle, G. and Pierrefite-Carle, V.: Preventive Cancer Stem Cell-Based Vaccination Reduces Liver Metastasis Development in a Rat Colon Carcinoma Syngeneic Model., Stem Cells 31, 423-32, 2013 |
| 大鼠 | Rat | 海马神经元 / Hippocampal neurons | Papain: 0.2% | Hibernate A | Mehta, N., Lopez, P., Vyas, A. and Schnaar, R.: Gangliosides and Nogo Receptors Independently Mediate Myelin-Associated Glycoprotein Inhibition of Neurite Outgrowth in Different Nerve Cells., J Biol Chem 282, 27875-86, 2007 |
| 大鼠 | Rat, male 12 week | 海绵浸润细胞 / Sponge infiltrating cells | Collagenase Type 4: 0.15%； Deoxyribonuclease I: 0.02% |
RPMI 1640 | Sharma N, Luo J, Kirschmann DA, O'Malley Y, Robbins ME, Akporiaye ET, Lubaroff DM, Heidger PM, Hendrix MJ: A novel immunological model for the study of prostate cancer, Cancer Res 59, 2271-6, 1999 |
| 大鼠 | Rat (ACI/NMs X BUF/Mna) F1, male, 28 months, rat, (ACI/MNs) male, 8 weeks | 上皮 / Epthelial | Collagenase Type 3: 0.1% | Eagle's MEM Serum-free | Masuda, A., Ohtsuka, K., and Matsuyama, M.: Establishment of Functional Epithelial Cell Lines from a Rat Thyoma and a Rat Thymus, In Vitro Cell Dev Biol 26, 713, 1990 |
| 大鼠 | Rat | 卵黄囊瘤 / Yolk sac tumor | Trypsin: 0.01% | DMEM | Brennan, M., Oldberg, A., Hayman, E., and Ruoslahti, E.: Effect of a Proteoglycan Produced by Rat Tumor Cells on Their Adhesion to Fibronectin-Collagen Substrata, Cancer Res 43, 4302, 1983 |
| 大鼠 | Rat, SD, female | 上皮，癌症和肿瘤 / Epithelial, cancer and tumor | Collagenase: 0.1% | Eagles's MEM | Cohen, L.: Isolation and Characterization of a Serially Cultivated, Neoplastic, Epithelial Cell Line from the N-nitrosomethylurea Induced Rat Mammary Adenocarcinoma, In Vitro 18, 565, 1982 |
| 大鼠 | Rat | 肿瘤，胰岛 / Tumor, islet | Trypsin: 0.05% | Medium 199 | Gazdar, A., Chick, W., Oie, H., Sims, H., King, D., Weir, G., and Lauris, V.: Continuous, Clonal, Insulin-and Somatostatin-Secreting Cell Lines Established from a Transplantable Rat Islet Cell Tumor, Proc Natl Acad Sci U S A 77 (6), 3519, 1980 |
| 大鼠 | Rat | 腹水性肝癌 / Ascites hepatoma | Trypsin: 0.1% | Phosphate buffer (see reference) | Essner, E.: Experiments on an Ascites Hepatoma. I. Enzymatic Digestion and Alkaline Degradation of the Cementing Substance and Separation of Cells, in Tumor Islands, Exp Cell Res 7, 430, 1954 |
