乳腺组织消化手册

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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。

乳腺组织消化手册

物种	详细来源	细胞名称	消化用酶	培养基	参考文献
牛	牛, 泌乳期	Mammary epithelial	Collagenase Type 3: 400 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
			Hyaluronidase: 100 u/ml
			Deoxyribonuclease I: 2 u/ml
牛	牛, 小母牛, 200公斤	Mammary epithelial	Collagenase Type 2: 1%	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
			Hyaluronidase: 1%
			Deoxyribonuclease I: 0.03%
牛	牛	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 Biol27, 585, 1991
牛	牛	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	牛, 幼年的, 哺乳期, 雌性	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	牛, 牛奶场, 纯血统的,	Secretory	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
牛	(包括大鼠, 豪斯曼, 白化变种, 白色)	Mammary gland	Collagenase: 0.02 - 0.03%	HBSS or EBSS
山羊	山羊, 哺乳期, 1 个月	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	豚鼠, 怀孕的, 4-10 天	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
人类	人类	Mammary Epithelial	Collagenase: 200 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
人类	人类	Mammary Epithelial	Hyaluronidase: 100 u/ml	DMEM/F-12
人类	人类, 正常活检	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
人类	人类	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
人类	人类	Epithelial	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
人类	人类	Fibroblasts	Collagenase Type 1: 450 IU/ml	DMEM/Ham's F-12
人类	人类	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
人类	人类, 女性, 15-61 岁大	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, 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	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	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
小鼠	小鼠	Mammary tumor	Hyaluronidase: 0.020%	DMEM/F12
小鼠	小鼠, 10 周	Fibroblasts, carcinoma	Collagenase Type 4: 0.5%	DMEM	Cheng, N. and Lambert, D.: Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice., J Vis Exp 54, e2716, 2011
			Trypsin: 0.2%
			Deoxyribonuclease I: 0.004
			Hyaluronidase: 1,000 u/ml
小鼠	小鼠, 12 周	Mammary epithelial	Collagenase: 0.3%	see reference	Taddei Ilaria, Deugnier Marie-Ange, Faraldo Marisa M, Petit Valerie, Bouvard Daniel, Medina Daniel, Fsssler Reinhard, Thiery Jean Paul, Glukhova Marina:Beta1 integrin deletion from the basal compartment of the mammary epithelium affects stem cells, Nat Cell Biol 10, 716-22, 2008
			Hyaluronidase: 100 u/ml
			Trypsin: 0.25%
			Neutral Protease: 0.5%
			Deoxyribonuclease I: 0.01%
小鼠	小鼠, 11 周	Epithelial	Collagenase Type 3: 25 u/ml	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
			Hyaluronidase: 0.1%
			Protease XIV: 0.05%
			Deoxyribonuclease I: 0.04%
小鼠	小鼠	Mammary epithelial stem	Collagenase Type 3: 0.1%	DMEM	Boulanger CA, Smith GH: Reducing mammary cancer risk through premature stem cell senescence, Oncogene 20, 2264-72, 2001
			Hyaluronidase: 0.1%
			Pronase: 1.25%
			Deoxyribonuclease I: 0.2%
小鼠	小鼠, 白变种实验室老鼠	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 Physiol153, 381, 1992
小鼠	小鼠, 白变种实验室老鼠, 处女, 雌性, 4 个月	Adipocytes	Trypsin: 50 礸/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 Biol25 (5), 409, 1989
小鼠	小鼠 (白变种实验室老鼠Crgl), 成年的	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	小鼠, 白变种实验室老鼠, 雌性, 6-8 周	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	小鼠 (C3H/HeN), 雌性	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	小鼠, (白变种实验室老鼠/Crgl Me), 雌性, 怀孕的	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	小鼠, 雌性	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	小鼠 (NMuMG), 雌性, 2 months	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	小鼠, 白变种实验室老鼠Cr1, 雌性	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	小鼠 (白变种实验室老鼠fC3H)	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	小鼠 (白变种实验室老鼠fC3H)	Epithelial	Collagenase: 1.0%	HBSS
小鼠	小鼠, 怀孕的 8-12 天	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	小鼠 (白变种实验室老鼠Crgl), 处女, 雌性, 4-8 个月	Epithelial	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	小鼠 (白变种实验室老鼠Crgl), 处女, 雌性, 4-8 个月	Mammary gland	Collagenase Type 3: 0.1%	HBSS
小鼠	小鼠 (白变种实验室老鼠), 怀孕的, 60-80 天	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	小鼠, 白变种实验室老鼠fC3H/Crgl, 处女, 雌性	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 雌性白变种实验室老鼠fC3H Mice, Cancer Res 38, 2103, 1978
小鼠	小鼠, 白变种实验室老鼠Crgl, 雌性	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	(包括豚鼠)	Swiss 3T3	Collagenase Type 3: 0.2%	PBS CMF
小鼠	小鼠, (C3H/Crgl or 白变种实验室老鼠Crgl), 8-10 天怀孕的	Epithelial	Collagenase Type 3: 0.12%	HBSS	Emerman, J., Enami, J., Pika, 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	小鼠, 哺乳（期）	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	小鼠, 雌性, 早期妊娠小于10 天	Epithelial	Collagenase: 0.1%	Eagle's MEM	Moore, D., and Lasfargues, E.: Method for the Continuous C*tion of Mammary Epithelium, In Vitro 7, 21, 1971
小鼠	小鼠, CBA, 处女, 9/10 周	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	小鼠, 哺乳期, 14-18 天	Parenchymal	Collagenase: 0.33%	Kreb's buffer	Pika, 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	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
小鼠	小鼠, 怀孕的, 14-17 天	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	小鼠, adult, 1-10 天s 怀孕的	Epithelial	Collagenase: 0.02%	Simm's	Lasfargues, E.: C*tion 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
小鼠	小鼠, adult, 1-10 天s 怀孕的	Mammary	Collagenase: 0.02%	Simm's
大鼠	大鼠	Mammary epithelial	Collagenase Type 3: 0.35%	HBSS	Mei, N., McDaniel, L., Dobrovolsky, V., Guo, X., Shaddock, J., Mitstaedt, 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	大鼠, 雌性, 50 天	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	大鼠, 雌性	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 雌性 Rat Growth and 5-hydroxymethyl-2'-deoxyuridine in DNA, Toxicol Sci 66, 125, 2002
大鼠	SD大鼠,	Mammary fibroblasts	Collagenase Type 3: 0.2%	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
			Neutral Protease: 0.2%
			Deoxyribonuclease I: 0.01%
大鼠	SD大鼠, 50 天	Mammary epithelial	Collagenase Type 3: 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	SD大鼠, 50 天	Mammary epithelial	Neutral Protease: 0.2%	DMEM/F12
大鼠	大鼠	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	(包括人类)	Epithelial	Collagenase Type 1: 0.4%	DMEM
大鼠	SD大鼠, 雌性, 60-90 天	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	Lewis大鼠, 雌性, 90 天	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	Lewis大鼠, 雌性, 90 天	Mammary	Collagenase: 0.05%	Medium 199
大鼠	SD大鼠, 雌性, 55 天	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	大鼠, 50-60 天	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	大鼠, 食鱼动物 344, 处女, 80-100 天	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	大鼠, LEW, 处女, 雌性, 45-50 天	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	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, 近亲繁殖的LEW, 雌性, 50-60 天大	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	Wistar大鼠, 产后的13-18 天	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	SD大鼠, 产后的15-20 天	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	大鼠, 处女	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

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