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        worthington 腫瘤組織分離指南

        發布者:艾美捷科技    發布時間:2022-06-10     
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        原代細胞分離中,針對腫瘤涉及到腫瘤,羊膜間充質干細胞、腫瘤浸潤淋巴細胞、上皮成釉細胞瘤,類器官,腦腫瘤和胰腺癌等等的解離通常會使用酶,如何選擇合適的酶來完成對應腫瘤的解離,以及原代細胞采集的常規操作, 艾美捷針對腫瘤相關的組織分類解離分種屬、不同的細胞、酶組合推薦,推薦濃度、培養基以及參考文獻,詳細匯總指南:腫瘤組織分離指南.png

         

        本技術指南描述了標準實驗室程序; 為建立細胞分離方案提供了一種合乎邏輯的實驗方法; 并列出了許多組織特異性參考資料,以幫助開發有效的方法。更多其他組織的解離指南,點擊查看:各組織分離指南。

         

        干細胞
        種屬種屬詳情細胞名稱酶組合推薦,含推薦濃度培養基參考文獻
        倉鼠Hamster, 90-100 g腫瘤 / TumorHyaluronidase: 0.1%Waymouth's MBGonzalez, 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
        倉鼠Hamster, 6 week old頰袋 / Buccal pouchNeutral Protease: 0.24%CMF HBSSMin, 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
        Human腫瘤浸潤淋巴細胞 / Tumor infiltrating lympocytesCollagenase Type 1: 200 u/mlRPMIJeske, 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
        Human腫瘤 / TumorCollagenase Type 4: 200 u/mlDMEMPhan, 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
        Human成纖維細胞 / FibroblastsCollagenase Type 1: 0.5%;
        Neutral Protease: 0.1%;
        Deoxyribonuclease I: 1,000 u/ml
        FM Fibroblast mediaAyuso, 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

        類器官 / OrganoidsCollagenase Type 4: 200-300 u/mlDMEMNeal, 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
        Human上皮成釉細胞瘤 / Ameloblastoma epithelialCollagenase: 0.1%;
        Neutral Protease: 50 u/ml
        MEMJiang, 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
        Human, 25-81 yr腦腫瘤 / Brain tumorNeutral Protease: 0.11 u/ml;
        Collagenase Type 4: 0.05%;
        Hyaluronidase: 1,000 u/ml;
        Deoxyribonuclease I: 5 u/ml
        HBSSVolovitz, 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
        Human胰腺癌 / Pancreatic cancerStemxyme 1: 0.2%HBSSPham, 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
        Human非小細胞肺腫瘤 / Non-small cell lung tumorCollagenase Type 1: 0.1%;
        Collagenase Type 2: 0.25%
        RPMI-1640Liang, 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
        Human肺腫瘤 / 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腫瘤浸潤淋巴細胞 / Tumour infiltrating lymphocytesCollagenase: 1%;
        Hyaluronidase: 1%;
        Deoxyribonuclease I: 3,000 u/ml
        RPMI 1640Baldan, 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
        Human結腸 / ColonTrypsin: 0.1%;
        Collagenase: 0.1%
        HBSSAli, 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
        Human膠質母細胞瘤 / Glioblastoma tumorTrypsin: 0.025%;
        Collagenase Type 4: 500 u/ml
        HBSSHasselbach, 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
        Human膠質瘤癌干細胞 / Glioma cancer stemPDS kit: per instructionsNeurobasalYan, 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
        Human黑色素瘤 / MelanomaCollagenase Type 4: 0.1%;
        Deoxyribonuclease I: 0.01%
        PBSWelte, 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
        Human腫瘤 / TumorCollagenase Type 1: 0.1%;
        Hyaluronidase: 0.01%;
        Deoxyribonuclease I: 0.01%
        DMEM/F12Chou, 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
        Human癌干細胞 / Cancer stemTrypsin: 0.25%;
        Collagenase: 0.05%;
        Hyaluronidase: 0.005%
        RPMIDobbin, 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
        Human大腸癌 / Colorectal cancerCollagenase Type 4: 2%;
        Deoxyribonuclease I: 0.1%
        RPMI 1640Zhou, 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
        Human腦腫瘤 / Brain tumor

        Collagenase Type 1: 0.04%;

        Hyaluronidase: 0.01%;

        Deoxyribonuclease I: 0.02%;

        Neutral Protease: 0.008%;

        DMEM/F12Hussein, 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
        Human致瘤性黑色素瘤 / Tumorigenic melanomaCollagenase Type 4: 200 u/ml;
        Trypsin: 0.05%;
        Deoxyribonuclease I: 50-100 u/ml
        PBSQuintana, 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
        Human胰腺腫瘤 / Pancreatic tumorCollagenase Type 4: 200 u/mlRPMI-1640Kim, 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
        Human腫瘤 / TumorCollagenase Type 4: 0.1%;
        Hyaluronidase: 0.07%;
        Deoxyribonuclease I: 0.04%
        see referenceSauvageot, 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
        Human結腸癌 / Colon cancerCollagenase Type 1: 300 u/ml;
        Hyaluronidase: 100 u/ml
        DMEM/F12Varnat, 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
        Human乳腺上皮 / Breast epithelialCollagenase Type 3: 200 u/mlHBSSLiu, 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


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