乳腺組織消化手冊(cè)

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Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年，但是對(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒有全面的理解。由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過去的經(jīng)驗(yàn)，而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案，以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。

細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞，而影響結(jié)果的原因有很多，包括（不限于）以下：

1、組織類型

2、物種來源

3、動(dòng)物年齡

4、遺傳改造情況（如knockouts）

5、所用的消化培養(yǎng)液

6、所用的酶

7、消化酶中的雜質(zhì)

8、所用消化酶的工作濃度

9、溫度

10、孵育時(shí)間

通常前四個(gè)原因沒辦法選擇，而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。

通過查閱文獻(xiàn)來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性，以及過去常用的粗制、未提純的組織分離酶。另外，組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類和糖脂組成，這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育（年齡）之間存在明顯的差異，這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶，如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。

本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識(shí)和方法，描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟，以便于制定一個(gè)*的細(xì)胞分離方案，同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。

下表列出多種組織的適宜消化酶（含參考文獻(xiàn)，以組織類型和物種分類，詳見鏈接）

脂肪組織	腎上腺組織	骨組織	腦組織
軟骨組織	結(jié)腸組織	內(nèi)皮組織	上皮組織
眼	心臟	腸	腎
肝	肺	淋巴結(jié)	乳腺
其它組織	其他組織	肌組織	神經(jīng)組織
胰腺	腮腺	垂體	前列腺
生殖	動(dòng)物鱗片	皮膚	脾臟
干細(xì)胞	胸腺	甲狀腺/甲狀旁腺	扁桃體
腫瘤

注：使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列，一般而言，Worthington公司的組織消化酶可用于大多組織類型的消化。

乳腺組織消化手冊(cè)

物種	詳細(xì)來源	細(xì)胞名稱	消化用酶	培養(yǎng)基	參考文獻(xiàn)
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, 牛奶場(chǎng), 純血統(tǒng)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, 白化變種, 白色)	Mammary gland	Collagenase: 0.02 - 0.03%	HBSS or EBSS
山羊	山羊, 哺乳期, 1 個(gè)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, 懷孕的, 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
人類	人類	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
人類	人類	Mammary Epithelial	Hyaluronidase: 100 u/ml	DMEM/F-12
人類	人類, 正常活檢	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
人類	人類	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
人類	人類	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
人類	人類	Fibroblasts	Collagenase Type 1: 450 IU/ml	DMEM/Ham's F-12
人類	人類	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
人類	人類, 女性, 15-61 歲大	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
人類	人類	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
人類	人類	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
人類	人類	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, 白變種實(shí)驗(yàn)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, 白變種實(shí)驗(yàn)室老鼠, 處女, 雌性, 4 個(gè)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 (白變種實(shí)驗(yàn)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, 白變種實(shí)驗(yàn)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, (白變種實(shí)驗(yàn)室老鼠/Crgl Me), 雌性, 懷孕的	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, 白變種實(shí)驗(yàn)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 (白變種實(shí)驗(yàn)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 (白變種實(shí)驗(yàn)室老鼠fC3H)	Epithelial	Collagenase: 1.0%	HBSS
小鼠	小鼠, 懷孕的 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 (白變種實(shí)驗(yàn)室老鼠Crgl), 處女, 雌性, 4-8 個(gè)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 (白變種實(shí)驗(yàn)室老鼠Crgl), 處女, 雌性, 4-8 個(gè)月	Mammary gland	Collagenase Type 3: 0.1%	HBSS
小鼠	小鼠 (白變種實(shí)驗(yàn)室老鼠), 懷孕的, 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, 白變種實(shí)驗(yàn)室老鼠fC3H/Crgl, 處女, 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 雌性白變種實(shí)驗(yàn)室老鼠fC3H Mice, Cancer Res 38, 2103, 1978
小鼠	小鼠, 白變種實(shí)驗(yàn)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 白變種實(shí)驗(yàn)室老鼠Crgl), 8-10 天懷孕的	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, 處女, 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, 懷孕的, 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 懷孕的	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 懷孕的	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	(包括人類)	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, 食魚動(dòng)物 344, 處女, 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, 處女, 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, 近親繁殖的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, 產(chǎn)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, 產(chǎn)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, 處女	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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