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：
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。
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| 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 |
| 小鼠 | Mouse (BALB/cCrgl), virgin, female, 4-8 month | 上皮，乳腺 / Epithelial，Mammary gland | Collagenase Type 3: 0.1% | HBSS | White, M., Hu, A., Hamamoto, S. and Nandi: In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: fibroblast-free growth and serial passage, In Vitro 14, 271, 1978 |
| 小鼠 | Mouse (BALB/c), pregnant, 60-80 days | 乳腺 / Mammary | Hyaluronidase: 0.1% | CMF medium | Ceriani, R., Peterson, J., and Abraham, S.: Removal of Cell Surface Material by Enzymes Used to Dissociate Mammary Gland Cells, In Vitro 14, 887, 1978 |
| 小鼠 | Mouse, BALB/cfC3H/Crgl, virgin, female | 结节转化 / Nodule-transformed | Hyaluronidase: 0.1% | Medium 199 | DeOme, K., Miyamoto, M., Osborn, R., Guzman, R., and Lum, K.: Detection of Inaparent Nodule-transformed Cells in the Mammary Gland Tissues of Virgn Female BALB/cfC3H Mice, Cancer Res 38, 2103, 1978 |
| 小鼠 | Mouse, BALB/cCrgl, female | 瑞士3T3 / Swiss 3T3 | Collagenase Type 3: 0.2% | PBS CMF | Asch, B., and Medina, D.: Concanavalin A-Induced Agglutinability of Normal, Preneoplastic, and Neoplastic Mouse Mammary Cells, J Natl Cancer Inst 61 (6), 1423, 1978 |
| 小鼠 | Mouse, (C3H/Crgl or BALB/cCrgl), 8-10 day pregnant | 上皮 / Epithelial | Collagenase Type 3: 0.12% | HBSS | Emerman, J., Enami, J., Pitelka, D., and Nandi, S.: Hormonal Effects on Intracellular and Secreted Casein in Cultures of Mouse Mammary Epithelial Cells on Floating Collagen Membranes, Proc Natl Acad Sci U S A 74 (10), 4466, 1977 |
| 小鼠 | Mouse, lactating | 实质细胞 / Parenchymal | Collagenase Type 1: 0.3% | Kreb's Ringer bicarbonate buffer | Kerkof, P., and Abraham, S.: Preparation of Adipose Cell-Free Suspensions of Mammary Gland Parenchymal Cells from Lactating Mice, Methods Enzymol 69, 693, 1976 |
| 小鼠 | Mouse, female, early pregnancy <10 days | 上皮 / Epithelial | Collagenase: 0.1% | Eagle's MEM | Moore, D., and Lasfargues, E.: Method for the Continuous Cultivation of Mammary Epithelium, In Vitro 7, 21, 1971 |
| 小鼠 | Mouse, CBA, virgin, 9/10 wk | 乳腺 / Mammary | Hyaluronidase: 0.1% | BSS, CMF | Prop, F., and Wiepjes, G.: Improved method for preparation of single-cell suspensions from mammary glands of adult virgin mouse, Exp Cell Res 61, 451, 1970 |
| 小鼠 | Mouse, lactating, 14-18 day | 实质细胞 / Parenchymal | Collagenase: 0.33% | Kreb's buffer | Pitelka, D., Kerkof, P., Gagne, H., Smith, S., and Abraham, S.: Characteristics of Cells Dissociated from Mouse Mammary Glands. I. Method of Separation and Morphology of Parenchymal Cells from Lactating Glands, Exp Cell Res 57, 43, 1969 |
| 小鼠 | Mouse, lactating, 14 day | 乳腺 / 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 |
| 小鼠 | Mouse, pregnant, 14-17 day | 乳腺 / Mammary | Collagenase: 0.05% - 0.1% | HBSS | Daniel, C., and DeOme, K.: Growth of Mouse Mammary Glands In Vivo After Monolayer Culture, Science 149, 634, 1965 |
| 小鼠 | Mouse, adult, 1-10 days pregnant | 上皮 / Epithelial | Collagenase: 0.02% | Simm's | Lasfargues, E.: Cultivation and Behavior In Vitro of the Normal Mammary Epithelium of the Adult Mouse. II. Observations on the Secretory Activity, Exp Cell Res 13, 553, 1957 |
| 大鼠 | Rat | 乳腺上皮 / Mammary epithelial | Collagenase Type 3: 0.35% | HBSS | Mei, N., McDaniel, L., Dobrovolsky, V., Guo, X., Shaddock, J., Mittelstaedt, R., Azuma, M., Shelton, S., McGarrity, L., Doerge, D. and Heflich, R.: The Genotoxicity of Acrylamide and Glycidamide in Big Blue Rats., Toxicol Sci 115, 412, 2010 |
| 大鼠 | Rat, female, 50 day | 乳腺上皮 / Mammary epithelial | Collagenase Type 3: 0.15% | DMEM/F12 | Maffini M., Soto A., Calabro J., Ucci A., and Sonnenschein C.: The Stroma as a Crucial Target in Rat Mammary Gland Carcinogenesis, J Cell Sci 117, 1495, 2004 |
| 大鼠 | Rat, female | 乳腺上皮 / Mammary gland epithelial | Collagenase Type 3: 0.35% | see reference | Djuric Z., Lewis S., Lu M., Mayhugh M., Naegeli L., Tang N., and Hart R.: Effect of Varying Caloric Restriction Levels on Female Rat Growth and 5-hydroxymethyl-2'-deoxyuridine in DNA, Toxicol Sci 66, 125, 2002 |
| 大鼠 | Rat, SD | 乳腺成纤维细胞 / Mammary fibroblasts | Collagenase Type 3: 0.2%； Neutral Protease: 0.2%； Deoxyribonuclease I: 0.01% |
DMEM/F-12 | Brake P., Zhang L., and Jefcoate C.: Aryl Hydrocarbon Receptor Regulation of Cytochrome P4501B1 in Rat Mammary Fibroblasts: Evidence for Transcriptional Repression by Glucocorticoids, Mol Pharmacol 54, 825, 1998 |
| 大鼠 | Rat, SD, 50 day | 乳腺上皮 / Mammary epithelial | Collagenase Type 3: 0.2%； Neutral Protease: 0.2% |
DMEM/F12 | Varela L., Darcy K., and Ip M.: The Epidermal Growth Factor Receptor is not Required for Tumor Necrosis Factor-alpha Action in Normal Mammary Epithelial Cells, Endocrinology 138, 3891, 1997 |
| 大鼠 | Rat | 上皮 / Epithelial | Collagenase Type 1: 0.4% | DMEM | Soriano, J., Pepper, M., Nakamura, T., Orci, L., and Montesano, R.: Hepatocyte Growth Factor Stimulates Extensive Development of Branching Duct-like Structures by Cloned Mammary Gland Epithelial Cells, J Cell Sci 108, 413-430, 1995 |
| 大鼠 | Rat, SD, female, 60-90 day | 上皮 / Epithelial | Collagenase Type 3: 0.5% | EBSS | Laduca, J., and Sinha, D.: In Vitro Carcinogenesis of Mammary Epithelial Cells by N-Nitroso-N-Methylurea Using a Collagen Gel Matrix Culture, In Vitro Cell Dev Biol 29A, 789, 1993 |
| 大鼠 | Rat, Lewis, female, 90 days | 上皮 / Epithelial | Collagenase: 0.05% | Medium 199 | Lin, T., Hom, Y.K., Richards, J. and Nandi, S.: Effects of Antioxidants and Reduced Oxygen Tension on Rat Mammary Epithelial Cells in Culture, In Vitro Cell Dev Biol 27A, 191, 1991 |
| 大鼠 | Rat, SD, female, 55 day | 上皮 / Epithelial | Neutral Protease: 3 u/ml | Medium 199 | Ehmann, U., Osborn, R., Guzman, R., and Fajardo, L.: Cultured Proliferating Rat Mammary Epithelial Cells, In Vitro Cell Dev Biol 27, 749, 1991 |
| 大鼠 | Rat, 50-60 days | 上皮 / Epithelial | Neutral Protease: 0.2% | EBSS | Hahm, H.A., Ip, M.M.: Primary Culture of Normal Rat Mammary Epithelial Cells Within a Basement Matrix. 1. Regulation of Proliferation by Hormones and Growth Factors, In Vitro Cell Dev Biol 26, 791, 1990 |
| 大鼠 | Rat, Fisher 344, virgin, 80-100 day | 上皮 / Epithelial | Hyaluronidase: 0.1% | Medium 199 | McGrath, M., Palmer, S., and Nandi, S.: Differential Response of Normal Rat Mammary Epithelial Cells to Mammogenic Hormones and EGF, J Cell Physiol 125, 182, 1985 |
| 大鼠 | Rat, LEW, virgin, female, 45-50 day | 上皮 / Epithelial | Collagenase: 0.1% | Medium 199 | Ethler, S.: Primary Culture and Serial Passage of Normal and Carcinogen-Treated Rat Mammary Epithelial Cells In Vitro, J Natl Cancer Inst 74 (6), 1307, 1985 |
| 大鼠 | Rat, SD, female | 上皮，癌症和肿瘤 / Epithelial, cancer and tumor | Collagenase: 0.1% | Eagles's MEM | Cohen, L.: Isolation and Characterization of a Serially Cultivated, Neoplastic, Epithelial Cell Line from the N-nitrosomethylurea Induced Rat Mammary Adenocarcinoma, In Vitro 18, 565, 1982 |
| 大鼠 | Rat, inbred LEW, female, 50-60 day old | 上皮 / Epithelial | Collagenase Type 3: 0.1% | Medium 199 | Richards, J., and Nandi, S.: Primary Culture of Rat Mammary Epithelial Cells. I. Effect of Plating Density, Horomones, and Serum on DNA Synthesis, J Natl Cancer Inst 61 (3), 765, 1978 |
| 大鼠 | Rat, Wistar, 13-18 day postpartum | 腺泡 / Acini | Collagenase: 0.05% | HBSS | Katz, J., Wals, P. and Van de Velde, R: Lipogenesis by Acini from Mammary Gland of Lactating Rats, J Biol Chem 249, 7348, 1974 |
| 大鼠 | Rat, SD, 15-20 day postpartum | 乳腺 / Mammary | Collagenase: 0.2% | Kreb's Ringer bicarbonate buffer | Martin, R., and Baldwin, R.: Effects of Insulin on Isolated Rat Mammary Cell Metabolism: Glucose Utilization and Metabolite Patterns, Endocrinology 89, 1263, 1971 |
| 大鼠 | Rat, virgin | 乳腺 / Mammary | Collagenase: 0.35% | Medium 199 | Moon, R., Janns, D., and Young, S.: Preparation of Fat Cell-"Free" Rat Mammary Gland, J Histochem Cytochem 17 (3), 182, 1969 |
