Worthington 乳腺组织分离指南

发布者：艾美捷科技发布时间：2022-06-07

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原代细胞分离中，针对乳腺组织涉及到乳腺上皮，上皮、乳腺、肌成纤维细胞等等的解离通常会使用酶，如何选择合适的酶来完成对应乳腺组织的解离，以及原代细胞采集的常规操作， 艾美捷针对乳腺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：乳腺组织分离指南

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。

乳腺
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
牛	Bovine, lactating Holstein	乳腺上皮 / Mammary epithelial	Collagenase Type 3: 400 u/ml； Hyaluronidase: 100 u/ml； Deoxyribonuclease I: 2 u/ml	HBSS	Miranda, S., Wang, Y., Purdie, N., Osborne, V., Coomber, B. and Cant, J.: Selenomethionine Stimulates Expression of Glutathione Peroxidase 1 and 3 and Growth of Bovine Mammary Epithelial Cells in Primary Culture., J Dairy Sci 92, 2670, 2009
牛	Bovine, heifer, 200kg	乳腺上皮 / Mammary epithelial	Collagenase Type 2: 1%； Hyaluronidase: 1%； Deoxyribonuclease I: 0.03%	M-199	Weber M., Purup S., Vestergaard M., Ellis S., Scndergard-Andersen J., Akers R., and Sejrsen K.: Contribution of Insulin-like Growth Factor (IGF)-I and IGF-binding Protein-3 to Mitogenic Activity in Bovine Mammary Extracts and Serum, J Endocrinol 161, 365, 1999
牛	Bovine	上皮 / Epithelial	Hyaluronidase: 0.005%	HBSS	Gibson, C., Vega, J., Baumrucker, C., Oakley, C., and Welsch, C.: Establishment And Characterization Of Bovine Mammary Epithelial Cell Lines, In Vitro Cell Dev Biol 27, 585, 1991
牛	Bovine	上皮 / Epithelial	Deoxyribonuclease I: 0.04%	HBSS/Medium 199	Baumrucker, C., Deemer, K., Walsh, R., Riss, T., and Akers, R.: Primary Culture of Bovine Mammary Acini on a Collagen Matrix, Tissue Cell 20 (4), 541, 1988
牛	Bovine, young, lactating, female	乳腺 / Mammary	Collagenase: 0.30 %	HBSS or EBSS	Anderson, C., and Larson, B.: Comparative Maintenance of Function in Dispersed Cell and Organ Cultures, Exp Cell Res 61, 24, 1970
牛	Bovine, dairy, purebred,	分泌，乳腺 / Secretory，Mammary gland	Collagenase: 0.02 - 0.03%	HBSS or EBSS	Schingoethe, D., Hageman, E., and Larson, B.: Essential Amino Acids for Milk Protein Synthesis in the In Vitro Secretory Cell and Stimulation by Elevated Levels, Biochim Biophys Acta 148, 469, 1967
山羊	Goat, lactating, 1 month	乳腺 / Mammary gland	Collagenase: 0.02 - 0.03%	HBSS or EBSS	Blanco, A., Rife, U., and Larson, B.: Lactate Dehydrogenase Isozymes during Dedifferentiation in Cultures of Mammary Secretory Cells, Nature 214, 1331, 1967
豚鼠	Guinea-pig, pregnant, 4-10 day	乳腺 / Mammary gland	Trypsin NF 1:250: 0.25%	Dulbecco phosphate	Turba, F., and Hilpert, N.: Secretion and Resorption of Proteins by Isolated Mammary Gland Cells. German, Biochem Z 334, 501, 1961
人	Human	乳腺上皮 / Mammary epithelial	Collagenase: 200 u/ml； Hyaluronidase: 100 u/ml	DMEM/F-12	Labarge, M., Garbe, J. and Stampfer, M.: Processing of Human Reduction Mammoplasty and Mastectomy Tissues for Cell Culture., J Vis Exp 71, , 2013
人	Human, normal biopsy	成纤维细胞 / Fibroblasts	Collagenase Type 3: 900 u/ml	DMEM/F-12	Ronnov-Jessen L., Villadsen R., Edwards J., and Petersen O.: Differential Expression of a Chloride Intracellular Channel Gene, CLIC4, in Transforming Growth Factor-beta1-mediated Conversion of Fibroblasts to Myofibroblasts, Am J Pathol 161, 471, 2002
人	Human	乳腺上皮细胞 / Mammary epithelial cells	Collagenase Type 1: 0.1%	DMEM	Huss, F. and Kratz, G.: Mammary Epithelial Cell and Adipocyte Co-Culture in a 3-D Matrix: The First Step Towards Tissue-Engineered Human Breast Tissue., Cells Tissues Organs 169, 361-7, 2001
人	Human	上皮细胞，成纤维细胞 / Epithelial，Fibroblasts	Collagenase Type 1: 450 IU/ml	DMEM/Ham's F-12	Ogmundsdottir, H., Petursdottir, I., Gudmundsdottir, I., Amundadottir, L., Ronnov-Jessen, L., and Petersen, O.: Effects of Lymphocytes and Fibroblasts on the Growth of Human Mammary Carcinoma Cells Studied in Short-Term Primary Cultures, In Vitro Cell Dev Biol 29A, 936, 1993
人	Human	肌成纤维细胞 / Myofibroblasts	Collagenase: 900 IU/ml	DME - F12	Ronnov-Jessen, L., VanDeurs, B., Nielsen, M., and Petersen, O.W.: Identification, Paracrine Generation, and Possible Function of Human Breast Carcinoma Myofibroblasts in Culture, In Vitro Cell Dev Biol 28, 273, 1992
人	Human, female, 15-61 yrs old	上皮 / Epithelial	Hyaluronidase: 150 IU/ml	DMEM	Berthon, P., Pancino, G., Cremoux, P., Roseto, A., Gespach, C., and Calvo, F.: Characterization of Normal Breast Epithelial Cells in Primary Cultures: Differentiation and Growth Factor Receptors Studies, J Tiss Cul Meth 28A, 716, 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，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	上皮 / Epithelial	Hyaluronidase: 100 u/ml	DMEM/Ham's F-12	Stampfer, M., Hallowes, R., and Hackett, A.: Growth of Normal Human Mammary Cells in Culture, In Vitro 16 (5), 415, 1980
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, 10 week	成纤维细胞，癌 / Fibroblasts, carcinoma	Collagenase Type 4: 0.5%； Trypsin: 0.2%； Deoxyribonuclease I: 0.004； Hyaluronidase: 1,000 u/ml	DMEM	Cheng, N. and Lambert, D.: Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice., J Vis Exp 54, e2716, 2011
小鼠	Mouse, 12 week	乳腺上皮 / Mammary epithelial	Collagenase: 0.3%； Hyaluronidase: 100 u/ml； Trypsin: 0.25%； Neutral Protease: 0.5%； Deoxyribonuclease I: 0.01%；	see reference	Taddei, I., Deugnier, M., Faraldo, M., Petit, V., Bouvard, D., Medina, D., Fassler, R., Thiery, J. and Glukhova, M.: Beta1 Integrin Deletion from the Basal Compartment of the Mammary Epithelium Affects Stem Cells, Nat Cell Biol 10, 716-22, 2008
小鼠	Mouse, 11 week	上皮 / Epithelial	Collagenase Type 3: 25 u/ml； Hyaluronidase: 0.1%； Protease XIV: 0.05%； Deoxyribonuclease I: 0.04%；	DMEM/F12	Mueller, S., Clark, J., Myers, P. and Korach, K.: Mammary Gland Development in Adult Mice Requires Epithelial and Stromal Estrogen Receptor Alpha., Endocrinology 143, 2357, 2002
小鼠	Mouse	乳腺上皮干细胞 / Mammary epithelial stem	Collagenase Type 3: 0.1%； Hyaluronidase: 0.1%； Pronase: 1.25%； Deoxyribonuclease I: 0.2%；	DMEM	Boulanger CA, Smith GH: Reducing mammary cancer risk through premature stem cell senescence, Oncogene 20, 2264-72, 2001
小鼠	Mouse, BALB/C	上皮 / Epithelium	Collagenase: 250 u/ml	HBSS	Kanazawa, T., and Hosick, H.: Transformed Growth Phenotype of Mouse Mammary Epithelium in Primary Culture Induced by Specific Fetal Mesenchymes, J Cell Physiol 153, 381, 1992
小鼠	Mouse, BALB/c, virgin, female, 4 month	脂肪细胞 / Adipocytes	Trypsin: 50 痢/ml	DMEM	Beck, J., Hosick, H., and Watkins, B.: Growth of Epithelium From a Preneoplastic Mammary Outgrowth on Response to Mammary Adipose Tissue, In Vitro Cell Dev Biol 25 (5), 409, 1989
小鼠	Mouse (BALB/cCrgl), mature	上皮 / Epithelial	Pronase: 0.01%	Hepes buffered medium 199	Bandyopadhyay, G., Imagawa, W., Wallace, D., and Nandi, S.: Proliferative effects of insulin and epidermal growth factor on mouse mammary epithelial cells in primary culture, J Biol Chem 263, 7567, 1988
小鼠	Mouse, BALB/c, female, 6-8 week	上皮 / Epithelial	Collagenase Type 3: 0.1%	DMEM	Ehmann, U., Peterson, W., and Misfeldt, D.: To Grow Mouse Mammary Epithelial Cells in Culture, J Cell Biol 98, 1026, 1984
小鼠	Mouse (C3H/HeN), female	上皮 / Epithelial	Deoxyribonuclease I: 0.0001%	Medium 199	Taketani, Y., and Oka, T.: EGF Stimulates Cell Proliferation and Inhibits Functional Differentiation of Mouse Mammalian Dipithelial Cells in Culture, Endocrinology 113, 871, 1983
小鼠	Mouse, (BALB/c/Crgl Me), female, pregnant	上皮 / Epithelial	Pepsin: 0.1% and 0.05%	HBSS with 0.2% EDTA, CMF	Riser, M., Huff, B., and Medina, D.: Pepsin Can Be Used To Subculture Viable Mammary Epithelial Cells, In Vitro 19, 730, 1983
小鼠	Mouse, female	上皮 / Epithelial	Deoxyribonuclease I: 0.1%	DMEM	Jones, W., and Hallowes, R.: Isolation of the Epithelial Subcomponents of the Mouse Mammary Gland for Tissue-Level Culture Studies, J Tiss Cul Meth 8 (1), 17, 1983
小鼠	Mouse (NMuMG), female, 2 months	上皮 / Epithelial	Collagenase Type 3: 0.1%	DMEM	Ehmann, U., and Misfeldt, D.: Mouse Mammary Cells in D-Valine Medium, In Vitro 18, 407, 1982
小鼠	Mouse, BALB/cCr1, female	上皮 / Epithelial	Collagenase Type 2: 0.2%	HBSS/DMEM	Asch, B., Burstein, N., Vidrich, A., and Sun, T.: Identification of Mouse Mammary Epithelial Cells by Immunofluorescence With Rabbit and Guinea-Pig Antikeratin Antisera, Proc Natl Acad Sci U S A 78, 5643, 1981
小鼠	Mouse (BALB/cfC3H)	乳腺肿瘤 / 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
小鼠	Mouse, pregnant 8-12 day	乳腺 / Mammary	(see reference):	HBSS	Yang J, Richards J, Guzman R, Imagawa W, and Nandi S.: Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gels, Proc Natl Acad Sci U S A 77, 2088, 1980