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：
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 / 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 |
| 倉鼠 | Hamster, 6 week old | 頰袋 / 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 | 腫瘤浸潤淋巴細胞 / 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 | 腫瘤 / 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 | 成纖維細胞 / 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 | 類器官 / 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 | 上皮成釉細胞瘤 / 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 | 腦腫瘤 / 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 | 非小細胞肺腫瘤 / 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 | 肺腫瘤 / 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 | 腫瘤浸潤淋巴細胞 / 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 | 結腸 / 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 | 膠質母細胞瘤 / 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 | 膠質瘤癌干細胞 / 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 | 腫瘤 / 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 | 癌干細胞 / 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 | 大腸癌 / 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 | 腦腫瘤 / 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 | 胰腺腫瘤 / 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 | 腫瘤 / 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 | 結腸癌 / 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 |
產品訂購：sales@amyjet.com
郵政編碼：430070
公司地址：武漢市洪山區光谷大道35號
光谷總部國際二期時代1棟13樓
版權所有：艾美捷科技有限公司 鄂ICP備10204150號-1 鄂公網安備：42018502004523號
微信掃碼在線客服
