worthington 肿瘤组织分离指南

发布者：艾美捷科技发布时间：2022-06-10

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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：肿瘤组织分离指南.png

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。

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