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Worthington组织消化指南
引言
组织消化酶主要用于组织培养和细胞生物学研究的原代细胞的分离和收集。尽管这些消化酶已经普遍应用多年，但是对其组织消化和细胞收集的作用机制却没有全面的理解。 由此导致的结果便是选择一种消化方法大多是随机选择或基于过去的经验，而不是在*理解消化原理的基础上选择的消化方案，以及如何根据具体实验目的修正消化方案以获得更好的实验结果。
细胞分离程序的目的是zui大程度的获得功能活性的单个细胞，而影响结果的原因有很多，包括（不限于）以下：
1、组织类型
2、物种来源
3、动物年龄
4、遗传改造情况（如knockouts）
5、所用的消化培养液
6、所用的酶
7、消化酶中的杂质
8、所用消化酶的工作浓度
9、温度
10、孵育时间
通常前四个原因没办法选择，而剩下的原因一般由经验决定以获取zui适实验结果。
通过查阅文献来选择*组织消化酶和zui适消化条件的研究人员通常被互相矛盾的实验数据搞得一头雾水。这些巨大差异来源于组织胞外基质的复杂而动态的特性，以及过去常用的粗制、未提纯的组织分离酶。另外，组织胞外基质由各种蛋白、糖蛋白、脂类和糖脂组成，这些成分的组成比例在不同物种、不同组织、甚至不同发育（年龄）之间存在明显的差异，这些因素都极大的影响了消化方案的制定。目前通常使用的粗制酶，如链霉蛋白酶、NF1:250、以及含有几种其他不同浓度蛋白酶的胶原酶、以及各种多糖酶、核酶和脂酶。
本指南总结了目前常用组织消化酶的zui适消化和细胞收集知识和方法，描述了标准的实验步骤， 以便于制定一个*的细胞分离方案，同时列举了一些相关的参考文献。
下表列出多种组织的适宜消化酶（含参考文献，以组织类型和物种分类，详见链接）
脂肪组织 | 肾上腺组织 | 骨组织 | 脑组织 |
软骨组织 | 结肠组织 | 内皮组织 | 上皮组织 |
眼 | 心脏 | 肠 | 肾 |
肝 | 肺 | 淋巴结 | 乳腺 |
其它组织 | 其他组织 | 肌组织 | 神经组织 |
胰腺 | 腮腺 | 垂体 | 前列腺 |
生殖 | 动物鳞片 | 皮肤 | 脾脏 |
干细胞 | 胸腺 | 甲状腺/甲状旁腺 | 扁桃体 |
肿瘤 |
|
|
|
注：使用Worthington公司消化酶的文献不限于上表链接中所列，一般而言，Worthington公司的组织消化酶可用于大多组织类型的消化。
肿瘤组织消化手册
| 物种 | 详细来源 | 细胞名称 | 消化用酶 | 培养基 | 参考文献 |
| 仓鼠 | 仓鼠, 90-100 克 | Tumor | Hyaluronidase: 0.1% | Waymouth's MB | Gonzalez, A., Oberley, T., Schultz, J., Ostrom, J., and Li, J.: In VitroCharactrization 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 | 仓鼠, 6 周大 | 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 |
| 人类 | 人类 | Non-small cell lung tumor | Collagenase Type 1: 0.1% | 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 |
| Collagenase Type 2: 0.25% | |||||
| 人类 | 人类 | Lung Tumor | Collagenase Type 1: 45-60 u/ml | 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 |
| Collagenase Type 2: 15-20 u/ml | |||||
| Collagenase Type 4: 45-60 u/ml | |||||
| Elastase: 0.002% | |||||
| Deoxyribonuclease I: 0.002% | |||||
| 人类 | 人类 | Glioblastoma Tumor | Trypsin: 0.025% | 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 |
| Collagenase Type 4: 500 u/ml | |||||
| 人类 | 人类 | Melanoma | Collagenase Type 4: 0.1% | 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 |
| Deoxyribonuclease I: 0.01% | |||||
| 人类 | 人类 | Tumor | Collagenase Type 1: 0.1% | 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 |
| Hyaluronidase: 0.01% | |||||
| Deoxyribonuclease I: 0.01% | |||||
| 人类 | 人类 | Colorectal cancer | Collagenase Type 4: 2% | 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 |
| Deoxyribonuclease I: 0.1% | |||||
| 人类 | 人类 | Tumorigenic melanoma | Collagenase Type 4: 200 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 |
| Trypsin: 0.05% | |||||
| Deoxyribonuclease I: 50-100 u/ml | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | Tumor | Collagenase Type 4: 0.1% | 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 |
| Hyaluronidase: 0.07% | |||||
| Deoxyribonuclease I: 0.04% | |||||
| 人类 | 人类 | Colon cancer | Collagenase Type 1: 300 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 |
| Hyaluronidase: 100 u/ml | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类, 成年人 | 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 |
| 人类 | 人类 | Colonocytes | Collagenase: | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类, 9-74 岁 | Neurofibroma | Neutral Protease: 1.25 u/ml | L-15 | Sheela S, Riccardi VM, Ratner N: Angiogenic and invasive properties of neurofibroma Schwann cells, J Cell Biol 111, 645-53, 1990 |
| Collagenase Type 1: 0.05% | |||||
| Hyaluronidase: 0.1% | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | Epthelial and tumor | 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 |
| Colon | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | Melanoma | 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 |
| Metastatic tumors | |||||
| 人类 | 人类 | Mammary tumors, hard | Collagenase: 0.10% | RPMI-1640 w/ 5% Fetal Calf Serum | Lasfargues, E.: Tissue Culture Methods/Applications,Kruse, P., and Patterson, M., Academic Press, 45, 1973 |
| 小鼠 | 小鼠 | Fibroblasts | Collagenase Type 2: 0.25% | 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 |
| Collagenase Type 4: 0.25% | |||||
| Deoxyribonuclease I: 0.05% | |||||
| 小鼠 | 小鼠 | 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 | 小鼠 | Mammary tumor | Collagenase: 0.15% | 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 |
| Hyaluronidase: 0.020% | |||||
| 小鼠 | 小鼠 | Rhabdomycosarcoma tumor | Trypsin: 0.012% | 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 Cell24, 710-24, 2013 |
| Collagenase Type 2: 0.1% | |||||
| 小鼠 | 小鼠 | 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 | 小鼠, 雄性 | Dendritic, macrophages | Collagenase (1 or 4): 100-200 u/ml | RPMI | Watkins, S., Zhu, Z., Watkins, K. and Hurwitz, A.: Isolation of Immune Cells from Primary Tumors., J Vis Exp 64, e3952, 2012 |
| Deoxyribonuclease I: 0.01% | |||||
| 小鼠 | 小鼠 | Lung tumor | Neutral Protease: 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 |
| Collagenase: 400 u/ml | |||||
| Deoxyribonuclease I: 50 u/ml | |||||
| 小鼠 | 小鼠 | 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 | 小鼠 | Pancreatic Cancer Stem | Collagenase Type 4: 200 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 |
| Neutral Protease: 0.6 u/ml | |||||
| 小鼠 | 小鼠 | Tumor infiltration lymphocytes | Collagenase Type 1: 0.25% | 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 |
| Collagenase Type 2: 0.15% | |||||
| Collagenase Type 4: 0.1% | |||||
| Hyaluronidase: 0.025% | |||||
| 小鼠 | 小鼠, 4-6 周 | 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 | 小鼠 | 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 | 小鼠, 30 周 | Tumor endothelial | Collagenase Type 4: 500 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 |
| Collagenase Type 2: 550 u/ml | |||||
| Deoxyribonuclease I: 3 u/ml | |||||
| 小鼠 | 小鼠 | Granule cell precursors, pre-neoplastic and tumor cells | Papain: 10 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 |
| Deoxyribonuclease I: 250 u/ml | |||||
| 小鼠 | 小鼠, 8-10 周 | 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 | 小鼠 | Pancreatic tumor | Collagenase Type 2: 0.5% | 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 |
| Collagenase Type 4: 0.5% | |||||
| Deoxyribonuclease I: 0.2% | |||||
| 小鼠 | 小鼠 | 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 | 小鼠, 雄性 | 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 | 小鼠 (BALB/cfC3H) | Mammary tumors | 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 |
| Epithelial | |||||
| 小鼠 | 小鼠, BALB/cfC3H/Crgl | Neoplastic | Trypsin: | DMEM | Hosick, H.: A Note on Growth Patterns of Epithelial Tumor Cells in Primary Culture, Cancer Res 34, 259, 1974 |
| Epithelial tumor | |||||
| 小鼠 | 小鼠, 哺乳（期）, 14 天 | 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 | 大鼠, 成年的 | Tumor | Collagenase Type 1: 0.16% | 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 |
| Hyaluronidase: 0.002% | |||||
| Deoxyribonuclease I: 0.006% | |||||
| 大鼠 | 大鼠, 雄性 12 周 | Sponge infiltrating cells | Collagenase Type 4: 0.15% | 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 |
| Deoxyribonuclease I: 0.02% | |||||
| 大鼠 | 大鼠 (ACI/NMs X BUF/Mna) F1, 雄性, 28 个月, 大鼠, (ACI/MNs) 雄性, 8 周 | 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 | 大鼠 | 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 | 大鼠, SD, 雌性 | Epithelial, cancer and tumor | Collagenase: 0.1% | Eagles's MEM | Cohen, L.: Isolation and Characterization of a Serially C*ted, Neoplastic, Epithelial Cell Line from the N-nitrosomethylurea Induced Rat Mammary Adenocarcinoma, In Vitro 18, 565, 1982 |
| 大鼠 | 大鼠 | 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 | 大鼠 | 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 |
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