脂肪組織消化手冊

更新時間：2016-02-17

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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和細胞生物學研究的原代細胞的分離和收集。盡管這些消化酶已經普遍應用多年，但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。由此導致的結果便是選擇一種消化方法大多是隨機選擇或基于過去的經驗，而不是在*理解消化原理的基礎上選擇的消化方案，以及如何根據具體實驗目的修正消化方案以獲得更好的實驗結果。

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

1、組織類型

2、物種來源

3、動物年齡

4、遺傳改造情況（如knockouts）

5、所用的消化培養液

6、所用的酶

7、消化酶中的雜質

8、所用消化酶的工作濃度

9、溫度

10、孵育時間

通常前四個原因沒辦法選擇，而剩下的原因一般由經驗決定以獲取zui適實驗結果。

通過查閱文獻來選擇*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驗數據搞得一頭霧水。這些巨大差異來源于組織胞外基質的復雜而動態的特性，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。

本指南總結了目前常用組織消化酶的zui適消化和細胞收集知識和方法，描述了標準的實驗步驟，以便于制定一個*的細胞分離方案，同時列舉了一些相關的參考文獻。

下表列出多種組織的適宜消化酶（含參考文獻，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	結腸組織	內皮組織	上皮組織
眼	心臟	腸	腎
肝	肺	淋巴結	乳腺
其它組織	其他組織	肌組織	神經組織
胰腺	腮腺	垂體	前列腺
生殖	動物鱗片	皮膚	脾臟
干細胞	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。

脂肪組織消化手冊

物種	詳細來源	細胞名稱	消化用酶	培養基	參考文獻
牛	牛	Adipocytes	Collagenase Type 1: 40 u/ml	Krebs-Ringer bicarbonate	Yang, Y., Baldwin, R.: Preparation and Metabolism of Isolated Cells from Bovine Adipose Tissue, J Dairy Sci 56, 350, 1973
犬	犬	Renal adipose derived cells	Collagenase Type 1: 0.3%	DMEM	Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011
犬	犬, 20-25公斤	Adipose stem cells	Collagenase: see reference	Media-199	Fischer, L., McIlhenny, S., Tulenko, T., Golesorkhi, N., Zhang, P., Larson, R., Lombardi, J., Shapiro, I. and DiMuzio, P.: Endothelial Differentiation of Adipose-Derived Stem Cells: Effects of Endothelial Cell Growth Supplement and Shear Force., J Surg Res 152, 157, 2009
犬	犬	White fat	Collagenase: 0.05%	Kreb's Ringer bicarbonate buffer	DiGirolamo, M., Mendlinger, S., and Fertig, J.W.: A Simple Method to Determine Fat Cell Size and Number in Four Mammalian Species, Am J Physiol 221, 850, 1971
馬	馬	Adipose derived stem cells	Collagenase Type 1: 0.1%	PBS	Vidal, M., Robinson, S., Lopez, M., Paulsen, D., Borkhsenious, O., Johnson, J., Moore, R. and Gimble, J.: Comparison of Chondrogenic Potential in Equine Mesenchymal Stromal Cells Derived from Adipose Tissue and Bone Marrow., Vet Surg 37, 713, 2008
馬	馬, 1-5 年	Adipose derived stem cells	Collagenase Type 1: 0.1%	PBS	Vidal, M., Kilroy, G., Lopez, M., Johnson, J., Moore, R. and Gimble, J.:Characterization of Equine Adipose Tissue-Derived Stromal Cells: Adipogenic and Osteogenic Capacity and Comparison with Bone Marrow-Derived Mesenchymal Stromal Cells., Vet Surg 36, 613, 2007
魚	魚, 大西洋鮭魚	Preadipocytes	Collagenase Type 1: 0.1%	HBSS	Todorcevic, M., Vegusdal, A., Gjoen, T., Sundvold, H., Torstensen, B., Kjaer, M. and Ruyter, B.: Changes in Fatty Acids Metabolism During Differentiation of Atlantic Salmon Preadipocytes; Effects of n-3 and n-9 Fatty Acids., Biochim Biophys Acta 1781, 326, 2008
沙鼠	不知年代的沙鼠	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer	Nedergaard, J. and Lindberg, O.: The Brown Fat Cell, Int Rev Cytol 74, 187, 1982
沙鼠	(還有小鼠, 倉鼠, 兔, 羔羊, 豚鼠)	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer
豚鼠	豚鼠, 成年的	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer	Nedergaard, J. and Lindberg, O.: The Brown Fat Cell, Int Rev Cytol 74, 187, 1982
豚鼠	(還有鼠, 倉鼠, 沙鼠, 兔, 羔羊)	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer
倉鼠	成年的倉鼠	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer	Nedergaard, J. and Lindberg, O.: The Brown Fat Cell, Int Rev Cytol 74, 187, 1982
倉鼠	(還有鼠, 沙鼠, 兔, 羔羊, 豚鼠)	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer
倉鼠	倉鼠, 5 周-12 個月	White fat	Collagenase: 0.05%	Kreb's Ringer bicarbonate buffer	DiGirolamo, M., Mendlinger, S., and Fertig, J.W.: A Simple Method to Determine Fat Cell Size and Number in Four Mammalian Species, Am J Physiol 221, 850, 1971
人	人	Mesenchymal Stromal Cells	Collagenase Type 4: 0.2%	PBS	Choudhery,M, Badowski, M, Muise, A and Harris, D: Effect of Mild Heat Stress on the Proliferative and Differentiative Ability of Human Mesenchymal Stromal Cells.,Cytotherapy 17, 359-68, 2015
人	人	Adipose derived stem cells	Collagenase Type 2: 0.1%	HBSS	Satish, L., Krill-Burger, J., Gallo, P., Etages, S., Liu, F., Philips, B., Ravuri, S., Marra, K., LaFramboise, W., Kathju, S. and Rubin, J.: Expression Analysis of Human Adipose-Derived Stem Cells During In Vitro Differentiation to an Adipocyte Lineage., BMC Med Genomics 8, 41, 2015
人	人, 女性	Adipose Stromal	Collagenase Type 1: 0.075%	DMEM	Kinoshita, K., Kuno, S., Ishimine, H., Aoi, N., Mineda, K., Kato, H., Doi, K., Kanayama, K., Feng, J., Mashiko, T., Kurisaki, A. and Yoshimura, K.:Therapeutic Potential of Adipose-Derived SSEA-3-Positive Muse Cells for Treating Diabetic Skin Ulcers., Stem Cells Transl Med 4, 146, 2015
人	人, 成年的	Adipose derived mesenchymal stem	Collagenase Type 2: 0.1%	see reference	Al-Saqi, S., Saliem, M., Asikainen, S., Quezada, H., Ekblad, A., Hovatta, O., Le Blanc, K., Jonasson, A. and Gotherstrom, C.: Defined Serum-Free Media for In Vitro Expansion of Adipose-Derived Mesenchymal Stem Cells., Cytotherapy 16, 915, 2014
人	人, 成年的	Adipose derived stem cells	Collagenase Type 1: 0.15%	DMEM	Koellensperger, E., Bollinger, N., Dexheimer, V., Gramley, F., Germann, G. and Leimer, U.: Choosing the Right Type of Serum for Different Applications of Human Adipose Tissue-Derived Stem Cells: Influence on Proliferation and Differentiation Abilities., Cytotherapy 16, 789, 2014
人	人	Mesenchymal Stromal Cells	Collagenase Type 1: 0.1%	PBS	Najar, M, Rodrigues, R, Buyl, K, Branson, S, Vanhaecke, T, Lagneaux, L, Rogiers, V and De Kock, J: Proliferative and Phenotypical Characteristics of Human Adipose Tissue-Derived Stem Cells: Comparison of Ficoll Gradient Centrifugation and Red Blood Cell Lysis Buffer Treatment Purification Methods.,Cytotherapy 16, 1220-8, 2014
人	人	Adipose stromal stem	Collagenase animal free: 200 u/ml	DMEM/Hams F-12	Carvalho, P., Gimble, J., Dias, I., Gomes, M. and Reis, R.: Xenofree Enzymatic Products for the Isolation of Human Adipose-Derived Stromal/Stem Cells., Tiss Eng19, 473-8, 2013
人	人	Stromal Vascular Fraction	Collagenase Type 1: 0.075%	PBS	Doi, K., Tanaka, S., Iida, H., Eto, H., Kato, H., Aoi, N., Kuno, S., Hirohi, T. and Yoshimura, K.: Stromal Vascular Fraction Isolated from Lipo-Aspirates Using an Automated Processing System: Bench and Bed Analysis., J Tissue Eng Regen Med 7, 864, 2013
人	人	Adipose-derived stem cells	Collagenase Type 1: 0.1%	PBS	Cervelli, V., Scioli, M., Gentile, P., Doldo, E., Bonanno, E., Spagnoli, L. and Orlandi, A.: Plaet-Rich Plasma Greatly Potentiates Insulin-Induced Adipogenic Differentiation of Human Adipose-Derived Stem Cells Through a Serine/Threonine Kinase Akt-dependent Mechanism and Promotes Clinical Fat Graft Maintenance.,Stem Cells Transl Med 1, 206-20, 2012
人	人	Perivascular	Collagenase Type 2: 0.1%	DMEM	James, A., Zara, J., Corselli, M., Askarinam, A., Zhou, A., Hourfar, A., Nguyen, A., Megerdichian, S., Asatrian, G., Pang, S., Stoker, D., Zhang, X., Wu, B., Ting, K., Peault, B. and Soo, C.: An Abundant Perivascular Source of Stem Cells for Bone Tissue Engineering., Stem Cells Transl Med 1, 673, 2012
人	人	Adipose derived stromal vascular	Collagenase Type 1: 0.1%	PBS	Gentile, P., Orlandi, A., Scioli, M., Di Pasquali, C., Bocchini, I. and Cervelli, V.:Concise Review: Adipose-Derived Stromal Vascular Fraction Cells and Plaet-Rich Plasma: Basic and Clinical Implications for Tissue Engineering Therapies in Regenerative Surgery., Stem Cells Transl Med 1, 230-6, 2012
人	人	Stromal vascular, adipocytes	Collagenase Type 1: 0.1%	DMEM	Gentile, P., Orlandi, A., Scioli, M., Di Pasquali, C., Bocchini, I., Curcio, C., Floris, M., Fiaschetti, V., Floris, R. and Cervell, V.: A Comparative Translational Study: The Combined Use of Enhanced Stromal Vascular Fraction and Plaet-Rich Plasma Improves Fat Grafting Maintenance in Breast Reconstruction., Stem Cells Transl Med 1, 341-51, 2012
人	人	Chorionic mesenchymal	Collagenase Type 1: 0.1%	MEM	Nazarov, I., Lee, J., Soupene, E., Etemad, S., Knapik, D., Green, W., Bashkirova, E., Fang, X., Matthay, M., Kuypers, F. and Serikov, V.: Multipotent Stromal Stem Cells from Human Placenta Demonstrate High Therapeutic Potential.,Stem Cells Transl Med 1, 359-72, 2012
			Neutral Protease: 2.5 u/ml
			Trypsin: 0.25%
人	人, 成年的	Stromovascular	Collagenase Type 1: 0.1%	PBS with BSA	Hagman, D., Kuzma, J., Larson, I., Foster-Schubert, K., Kuan, L., Cignarella, A., Geamanu, E., Makar, K., Gottlieb, J. and Kratz, M.: Characterizing and Quantifying Leukocyte Populations in Human Adipose Tissue: Impact of Enzymatic Tissue Processing., J Immunol Methods 386, 50, 2012
人	人	Adipose derived stromal	Collagenase Type 1: 0.1%	PBS	Naaijkens, B., Niessen, H., Prins, H., Krijnen, P., Kokhuis, T., de Jong, N,, van Hinsbergh, V., Kamp, O., Helder, M., Musters, R., van Dijk, A. and Juffermans, L.: Human Plaet Lysate as a Fetal Bovine Serum Substitute Improves Human Adipose-Derived Stromal Cell Culture for Future Cardiac Repair Applications., Cell Tissue Res 348, 119, 2012
人	人	Adipose derived stem	Collagenase Type 1: 0.1%	DMEM/F-12	Wu, I., Nahas, Z., Kimmerling, K., Rosson, G. and Elisseeff, J.: An Injectable Adipose Matrix for Soft-Tissue Reconstruction., Plast Reconstr Surg 129, 1247, 2012
人	人, 男性 40-60歲	Adipose derived stem cells	Collagenase: 0.25%	PBS	Blasi, A., Martino, C., Balducci, L., Saldarelli, M., Soleti, A., Navone, S., Canzi, L., Cristini, S., Invernici, G., Parati, E. and Alessandri, G.: Dermal Fibroblasts Display Similar Phenotypic and Differentiation Capacity to Fat-Derived Mesenchymal Stem Cells, but Differ in Anti-Inflammatory and Angiogenic Potential,Vasc Cell 3, 5, 2011
人	人, 男性 40-60歲	Adipose derived stem cells	Deoxyribonuclease I: 0.002%	PBS
人	人	Renal adipose derived cells	Collagenase Type 1: 0.3%	DMEM	Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011
人	人	Adipose derived mesenchymal	Collagenase Type 1: 0.1%	L-DMEM	Yang, X., He, X., He, J., Zhang, L., Su, X., Dong, Z., Xu, Y., Li, Y. and Li, YL.:High Efficient Isolation and Systematic Identification of Human Adipose-Derived Mesenchymal Stem Cells., J Biomed Sci 18, 59, 2011
人	人	Adipose derived stem	Collagenase Type 1: 0.1%	DMEM/Ham's F-12	Yu, G. Floyd, ZE., Wu, X., Halvorsen, Y. and Gimble, J.: Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates., Methods Mol Biol 702, 17-27, 2011
人	人	Adipocytes	Collagenase Type 1: 0.1%	RPMI	Basu, S., Haghiac, M., Surace, P., Challier, J., Guerre-Millo, M., Singh, K., Waters, T., Minium, J., Presley, L., Catalano, P. and Hauguel-de Mouzon, S.:Pregravid Obesity Associates with Increased Maternal Endotoxemia and Metabolic Inflammation., Obesity 19, 476, 2011
人	人	Adipocytes	Collagenase Type 1: 0.1%	KRB	Peters, R., Wolf, M., van den Broek, M., Nuvolone, M., Dannenmann, S., Stieger, B., Rapold, R., Konrad, D., Rubin, A., Bertino, J., Aguzzi, A., Heikenwalder, M. and Knuth, A.: Efficient Generation of Multipotent Mesenchymal Stem Cells from Umbilical Cord Blood in Stroma-Free Liquid Culture.,PLoS ONE 5, e15689, 2010
人	人	Adipose tissue-derived stem	Collagenase Type 2: 1.0%	DMEM/F12	Tan, H., DeFail, A., Rubin, J., Chu, C. and Marra, K.: Novel Multiarm PEG-Based Hydrogels for Tissue Engineering., J Biomed Mater Res 92, 979, 2010
人	人	Stromal vascular	Collagenase Type 2: 0.1%	HBSS	Zimmerlin, L., Donnenberg, V., Pfeifer, M., Meyer, EM, Peault, B., Rubin, JP and Donnenberg, A.: Stromal Vascular Progenitors in Adult Human Adipose Tissue.,Cytometry 77, 22-30, 2010
人	人	Adipose stromal	Collagenase Type 1: 0.1%	DMEM/Hams F-12	Yu, G., Wu, X., Dietrich, M., Polk, P., Scott, LK, Ptitsyn, A. and Gimble, J.: Yield and Characterization of Subcutaneous Human Adipose-Derived Stem Cells by Flow Cytometric and Adipogenic mRNA Analyzes., Cytotherapy 12, 538, 2010
人	人	Adipocytes	Collagenase: 0.075%	DMEM	Suga, H., Eto, H., Shigeura, T., Inoue, K., Aoi, N., Kato, H., Nishimura, S., Manabe, I., Gonda, K. and Yoshimura, K.: IFATS Collection: Fibroblast Growth Factor-2-Induced Hepatocyte Growth Factor Secretion by Adipose-Derived Stromal Cells Inhibits Postinjury Fibrogenesis Through a c-Jun N-Terminal Kinase-Dependent Mechanism., Stem Cells 27, 238, 2009
人	人	Adipose Stromal Cell	Collagenase Type 1: 0.1%	DMEM	Cai, L., Johnstone, B., Cook, T., Tan, J., Fishbein, M., Chen, P. and March, K.:IFATS Collection: Human Adipose Tissue-Derived Stem Cells Induce Angiogenesis and Nerve Sprouting Following Myocardial Infarction, in Conjunction with Potent Preservation of Cardiac Function., Stem Cells 27, 230, 2009
人	人	Adipose tissue-derived stem	Collagenase Type 2: 1.0%	DMEM/F12	Tan, H., Ramirez, C., Miljkovic, N., Li, H., Rubin, J. and Marra, K.:Thermosensitive Injectable Hyaluronic Acid Hydrogel for Adipose Tissue Engineering., Biomaterials 30, 6844, 2009
人	人	Adipose derived stem cells	Collagenase Type 1: 0.1%	DMEM	Tandon, N., Goh, B., Marsano, A., Chao, P., Montouri-Sorrentino, C., Gimble, J. and Vunjak-Novakovic, G.: Alignment and Elongation of Human Adipose-Derived Stem Cells in Response to Direct-Current Electrical Stimulation., Conf Proc IEEE Eng Med Biol Soc 2009, 6517-21, 2009
人	人	Adipose derived stromal cells	Collagenase Type 1: see reference	DMEM	Traktuev, D., Merfeld-Clauss, S., Li, J., Kolonin, M., Arap, W., Pasqualini, R., Johnstone, B., and March, K.: A Population of Multipotent CD34-Positive Adipose Stromal Cells Share Pericyte and Mesenchymal Surface Markers, Reside in a Periendothelial Location, and Stabilize Endothelial Networks, Circ Res 102, 77, 2008
人	人	Stromal	Collagenase Type 1: 0.2%	PBS	Minana, M., Carbonell-Uberos, F., Mirabet, V., Marin, S. and Encabo, A.: IFATS Collection: Identification of Hemangioblasts in the Adult Human Adipose Tissue.,Stem Cells 26, 2696, 2008
人	人	Stromal	Collagenase Type 1: 0.1%	DMEM	Nie, J., Chang, B., Traktuev, D., Sun, J., March, K., Chan, L., Sage, H., Pasqualini, R., Arap, W. and Kolonin, M.: IFATS Collection: Combinatorial Peptides Identify alpha-5 beta-1 Integrin as a Receptor for the Matricellular Protein SPARC on Adipose Stromal Cells., Stem Cells 26, 2735, 2008
人	人	Stromal	Neutral Protease:	DMEM
人	人	Stem Cells	Collagenase: 280 u/ml	D-PBS	Pilgaard, L., Lund, P., Rasmussen, J., Fink, T. and Zachar, V.: Comparative Analysis of Highly Defined Proteases for the Isolation of Adipose Tissue-Derived Stem Cells., Regen Med 3, 705-15, 2008
人	人	Adipocytes	Collagenase Type 1: 0.2%	HBSS	Bujalska Iwona J, Durrani Omar M, Abbott Joseph, Onyimba Claire U, Khosla Pamela, Moosavi Areeb H, Reuser Tristan T Q, Stewart Paul M, Tomlinson Jeremy W, Walker Elizabeth A, Rauz Saaeha: Characterisation of 11beta-hydroxysteroid dehydrogenase 1 in human orbital adipose tissue: a comparison with subcutaneous and omental fat, J Endocrinol 192, 279-88, 2007
人	人	Adipose derived stromal cells	Collagenase Type 1: 0.15%	DMEM	Schaffler A, Buchler C: Concise review: adipose tissue-derived stromal cells--basic and clinical implications for novel cell-based therapies, Stem Cells 25, 818-27, 2007
人	人	Adipose derived stem cells	Collagenase Type 1: 0.1%	DMEM/F12	Kilroy, G., Foster, S., Wu, X., Ruiz, J., Sherwood, S., Heifetz, A., Ludlow, J., Stricker, D., Potiny, S., Green, P., Halvorsen, Y., Cheatham, B., Storms, R. and Gimble, J.: Cytokine Profile of Human Adipose-Derived Stem Cells: Expression of Angiogenic, Hematopoietic, and Pro-Inflammatory Factors., J Cell Physiol 212, 702-9, 2007
人	人	Mesenchymal stem	Collagenase Type 1: 0.1%	HBSS	Jeon Eun Su, Song Hae Young, Kim Mi Ra, Moon Hyun Jung, Bae Yong Chan, Jung Jin Sup, Kim Jae Ho: Sphingosylphosphorylcholine induces proliferation of human adipose tissue-derived mesenchymal stem cells via activation of JNK, J Lipid Res 47, 653-64, 2006
人	人	Preadipocytes	Collagenase Type 1: 196 u/ml	M199	Koellensperger, E., Von Heimburg, D., Markowicz, M. and Pallua, N.: Human Serum from Plaet-Poor Plasma for the Culture of Primary Human Preadipocytes.,Stem Cells 24, 1218-25, 2006
人	人	Adipocytes, stromal vascular	Collagenase: 0.2%	HBSS	Boquest Andrew C, Shahdadfar Aboulghassem, Fronsdal Katrine, Sigurjonsson Olafur, Tunheim Siv H, Collas Philippe, Brinchmann Jan E:Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture, Mol Biol Cell 16, 1131-41, 2005
人	人	Multipotent adipose derived stem	Collagenase: 0.2%	DMEM	Rodriguez Anne-Marie, Pisani Didier, Dechesne Claude A, Turc-Carel Claude, Kurzenne Jean-Yves, Wdziekonski Brigitte, Villageois Albert, Bagnis Claude, Breittmayer Jean-Philippe, Groux Herve, Ailhaud Gerard, Dani Christian:Transplantation of a multipotent cell population from human adipose tissue induces dystrophin expression in the immunocompetent mdx mouse, J Exp Med 201, 1397-405, 2005
人	人	成年的 stem cells	Collagenase Type 1: 0.1%	PBS	Devireddy, R., Thirumala, S. and Gimble, J.: Cellular Response of Adipose Derived Passage-4 Adult Stem Cells to Freezing Stress., J Biomech Eng 127, 1081, 2005
人	人, 成年的 , 肥胖的	Adipocytes	Collagenase Type 2: 0.1%	DMEM/F12	Seboek, D., Linscheid, P., Zulewski, H., Langer, I., Christ-Crain, M., Keller, U., and Muller, B.: Somatostatin Is Expressed and Secreted by Human Adipose Tissue Upon Infection and Inflammation, J Clin Endocrinol Metab 89(10), 4833, 2004
人	人, 成年的, 女性	Preadipocytes	Collagenase Type 1: 0.2%	DMEM/F-12	Quickler, M., Sinha, B., Tomlinson, J., Bujalska, I., Stewart, P., Arlt, W.,:Androgen generation in adipose tissue in women with simple obesity--a site-specific role for 17beta-hydroxysteroid dehydrogenase type 5, J Endocrinol 183, 331, 2004
人	人,成年的	Adipocytes	Collagenase Type 1: 0.13%	see reference	Fain JN, Madan AK, Hiler ML, Cheema P, and Bahouth SW.: Comparison of the release of adipokines by adipose tissue, adipose tissue matrix, and adipocytes from visceral and subcutaneous abdominal adipose tissues of obese humans,Endocrinology 145, 2273, 2004
人	人	Stromal vascular, adipocytes	Collagenase: 0.2%	DMEM/F12	Planat-Benard Valerie, Silvestre Jean-Sebastien, Cousin Beatrice, Andre Mireille, Nibbelink Maryse, Tamarat Radia, Clergue Michel, Manneville Carole, Saillan-Barreau Corinne, Duriez Micheline, Tedgui Alain, Levy Bernard, Penicaud Luc, Casteilla Louis: Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives, Circulation 109, 656-63, 2004
人	人	Stromal vascular, adipocytes, stem	Collagenase: 300 u/ml	PBS	Miranville A, Heeschen C, Sengenes C, Curat CA, Busse R, Bouloumie A:Improvement of postnatal neovascularization by human adipose tissue-derived stem cells, Circulation 110, 349-55, 2004
人	人, 成年的, 女性, 非糖尿病的	Adipocytes	Collagenase Type 2: 0.05%	DMEM	Gesta, S., Lolmede, K., Daviaud, D., Berlan, M., Bouloumie, A., Lafontan, M., Valet, P., and Saulnier-Blache, J.: Culture of human adipose tissue explants leads to profound alteration of adipocyte gene expression, Horm Metab Res 35, 158, 2003
人	人	Adipocytes	Collagenase Type 1: see reference	Saline	Patwardhan, R., Tubbs, R., Leonard, R., Kelly, D., Killingsworth, C., Rollins, D., Smith, W., Ideker, R., and Oakes, W.: Discernment of Adipose versus Nervous Tissue: A Novel Adjunct Solution in Lipomyelomeningocele Surgery, Ped Neurosurg 36, 314, 2002
人	人, 成年的, 男性和女性	Adipocytes	Collagenase Type 1: 0.2%	DMEM/F12	McTernan, P., Anderson, L., Anwar, A., Eggo, M., Crocker, J., Barnett, A., Stewart, P., and Kumar, S.: Glucocorticoid Regulation of P450 Aromatase Activity in Human Adipose Tissue: Gender and Site Differences, J Clin Endocrinol Metab 87, 1327, 2002
人	人	Processed lipoaspi鼠e cells	Collagenase Type 1: 0.075%	PBS	Zuk Patricia A, Zhu Min, Ashjian Peter, De Ugarte Daniel A, Huang Jerry I, Mizuno Hiroshi, Alfonso Zeni C, Fraser John K, Benhaim Prosper, Hedrick Marc H: Human adipose tissue is a source of multipotent stem cells, Mol Biol Cell 13, 4279-95, 2002
人	人	Adipocytes	Collagenase Type 1: 0.2%	HBSS	McTernan PG, Anwar A, Eggo MC, Barnett AH, Stewart PM, Kumar S: Gender differences in the regulation of P450 aromatase expression and activity in human adipose tissue, Int J Obes Relat Metab Disord 24, 875-81, 2000
人	人, 成年的, 男性和女性	Adipocytes	Collagenase Type 1: 0.1%	DMEM/Ham's F-12	Gottschling-Zelle, H., Birgel, M., Scriba, D., Blum, W., and Hauner, H.: Depot-specific Release of Leptin from Subcutaneous and Omental Adipocytes in Suspension Culture: Effect of Tumor Necrosis Factor-alpha and Transforming Growth Factor-beta1, Eur J Endocrinol 141 (4), 436, 1999
人	人, 成年的, 男性和女性	Adipocytes	Collagenase Type 1: 0.1%	DMEM/F12	Zhang, H., Kumar, S., Barnett, A., and Eggo, M.: Intrinsic Site-Specific Differences in the Expression of Leptin in Human Adipocytes and Its Autocrine Effects on Glucose Uptake, J Clin Endocrinol Metab 84, 2550, 1999
人	人, 非糖尿病的, 男性	Adipocytes	Collagenase Type 1: 0.05%	Kreb's Ringer bicarbonate buffer	Anderson, O., Gliemann, J., and Gammeltoft: Receptor Binding and Biological Effect of Insulin in Human Adipocytes, Diabetologia 13, 589, 1977
小鼠	小鼠	Adipocytes	Collagenase: 0.1%	Krebs	Jang, H., Bhasin, S., Guarneri, T., Serra, C., Schneider, M., Lee, M., Guo, W., Fried, S., Pencina, K. and Jasuja, R.: The Effects of A Single Developmentally-Entrained Pulse of Testosterone in Female Neonatal Mice On Reproductive and Metabolic Functions in Adult Life., Endocrinology , EN20151117, 2015
小鼠	小鼠, 20 周	Stromal Vascular	Collagenase Type 2: 0.2%	RPMI-1640	Kondo, T, Toyoshima, Y., Ishii, Y and Kyuwa, S.: Natural Killer T Cells in Adipose Tissue are Activated in Lean Mice., Exp Anim 62, 319, 2013
小鼠	小鼠	Adipose derived stem	Collagenase Type 2: 0.1%	DMEM	Takahashi, H., Haraguchi, N., Nishikawa, S., Miyazaki, S., Suzuki, Y., Mizushima, T., Nishimura, J., Takemasa, I., Yamamoto, H., Mimori, K., Ishii, H., Doki, Y. and Mori, M.: Biological and Clinical Availability of Adipose-Derived Stem Cells for Pelvic Dead Space Repair., Stem Cells Transl Med 1, 803, 2012
小鼠	小鼠	Stem and progenitor	Collagenase Type 2: 0.2%	HBSS	Han, J., Koh, Y., Moon, H., Ryoo, H., Cho, C., Kim, I. and Koh, G.: Adipose Tissue is an Extramedullary Reservoir for Functional Hematopoietic Stem and Progenitor Cells., Blood 115, 957, 2010
小鼠	小鼠, 3 周	Adipocytes	Collagenase Type 1: 0.2%	HBSS	De Matteis, R., Zingaretti, M., Murano, I., Vitali, A., Frontini, A., Giannulis, I., Barbali, G., Marcucci, F., Bordicchia, M., Sarzani, R., Raviola, E. and Cinti, S.:In Vivo Physiological Transdifferentiation of Adult Adipose Cells., Stem Cells 27, 2761, 2009
小鼠	小鼠	White adipocytes	Collagenase Type 2: 0.1%	DMEM	Wong, K., Szeto, F, Zhang, W., Ye, H., Kong, J., Zhang, Z., Sun, X. and Li, Y:Involvement of the Vitamin D Receptor in Energy Metabolism: Regulation of Uncoupling Proteins., Am J Physiol/Endo 296, 820, 2009
小鼠	小鼠	Adipose Stromal Cell	Collagenase Type 2: 0.075%	DMEM	Thangarajah, H., Vial, I., Chang, E., El-Ftesi, S., Januszyk, M., Chang, E., Paterno, J., Neofytou, E., Longaker, M. and Gurtner, G.: IFATS Collection: Adipose Stromal Cells Adopt a Proangiogenic Phenotype Under the Influence of Hypoxia., Stem Cells 27, 266, 2009
小鼠	小鼠	Adipose tissue-derived stem	Collagenase Type 1: 0.075%	Modified Eagles	Cho, K., Park, H., Park, H., Jung, J., Jeon, S., Kim, Y. and Roh, H.: IFATS Collection: Immunomodulatory Effects of Adipose Tissue-Derived Stem Cells in an Allergic Rhinitis Mouse Model., Stem Cells 27, 259-65, 2009
小鼠	小鼠, C57Bl/6J	Adipocytes	Collagenase Type 1: 0.15%	DMEM/F12	Aoyagi T, Shimba S, Tezuka M: Characteristics of Circadian Gene Expressions in Mice White Adipose Tissue and 3T3-L1 Adipocytes, J Health Sci 51, 21, 2005
小鼠	小鼠, 3-6 天和 2-3 個月	Adipose-derived stromal cells	Collagenase Type 2: 0.075%	PBS	Cowan Catherine M, Shi Yun-Ying, Aalami Oliver O, Chou Yu-Fen, Mari Carina, Thomas Romy, Quarto Natalina, Contag Christopher H, Wu Benjamin, Longaker Michael T: Adipose-derived adult stromal cells heal critical-size mouse calvarial defects, Nat Biotechnol 22, 560-7, 2004
小鼠	小鼠	Stromal vascular, adipocytes	Collagenase: 0.2%	DMEM/F12	Planat-Benard Valerie, Silvestre Jean-Sebastien, Cousin Beatrice, Andre Mireille, Nibbelink Maryse, Tamarat Radia, Clergue Michel, Manneville Carole, Saillan-Barreau Corinne, Duriez Micheline, Tedgui Alain, Levy Bernard, Penicaud Luc, Casteilla Louis: Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives, Circulation 109, 656-63, 2004
小鼠	小鼠, C57BL/6J and FVB, 雄性, 8-9 周	Adipocytes	Collagenase: see reference	DMEM	Ruan, H., Zarnowski, MJ., Cushman, S., and Lodish, H.: Standared Isolation of Primary Adipose Cells from Mouse Epididymal Fat Pads Induces Inflammatory Mediators and Down-regulates Adipocyte Genes, J Biol Chem 278, 47585, 2003
小鼠	小鼠, 雌雄合計	Adipocytes	Collagenase: 0.05%	Krebs-Ringer Phosphate HEPES (KRPH)	Nadler, S., Stoehr, J., Rabaglia, M., Schueler, K., Birnbaum, M., and Attie, A.:Normal Akt/PKB with Reduced PI3K Activation in Insulin-resistant Mice, Am J Physiol/Endo 281, E1249, 2001
小鼠	小鼠, B6D2F1,雜交子代,新西蘭黑雌性 & 新西蘭黑雄性	Vascular endothelial	Collagenase: 0.2%	PBS	Launder, T., Gegen, N., Knedler, A., and Harbeck, R.: The Isolation and Characterization of Enriched Microvascular Endothelial Cells From Mouse Adipose Tissue, J Immunol Methods 102, 45, 1987
豬	豬, 雌性, 一年以下	Adipose mesenchymal stem	Collagenase Type 1: 0.1%	DMEM	Williams, K., Picou, A., Kish, S., Giraldo, A., Godke, R. and Bondioli, K: Isolation and Characterization of Porcine Adipose Tissue-Derived Adult Stem Cells., Cells Tissues Organs 188, 251, 2008
豬	豬, 雜交繁育, 雄性 1-4 天	Adipocytes	Collagenase Type 1: 0.2%	DMEM/F12	Ramsay, T.G.: Porcine Leptin Inhibits Lipogenesis in Porcine Adipocytes, J Anim Sci 81, 3008, 2003
豬	豬, 雜交繁育的	Adipocytes	Collagenase Type 1: 0.067%	Krebs-Ringer	Liang, W. and Mills, S.: Quantitative Analysis of beta-Adrenergic Receptor Subtypes in Pig Tissues., J Anim Sci 80, 963-70, 2002
豬	豬, 8-9 周	Adipose	Collagenase Type 1: 300 u/ml	HEPES	Ding, S., McNeel, R., and Mersmann, H.: Expression of Porcine Adipocyte Transcripts: Tissue Distribution and Differentiation In Vitro and In Vivo, Comp Biochem Physiol B 123, 307, 1999
豬	豬, 初生	Adipocytes	Collagenase Type 1: 0.3%	Krebs-Ringer bicarbonate albumin	Wang Y, Fried SK, Petersen RN, Schoknecht PA: Somatotropin regulates adipose tissue metabolism in neonatal swine, J Nutr 129, 139-45, 1999
豬	豬, 雜交繁育, 1-3 天	Adipose, Stromal-vascular	Collagenase Type 1: 0.2%	DMEM/F12	Suryawan, A., Swanson, L., and Hu, C.: Insulin and Hydrocortisone, But Not Triiodothyronine, Are Required for the Differentiation of Pig Preadipocytes in Primary Culture, J Anim Sci 75, 105, 1997
小鼠	小鼠	Mesenchymal Stem Cells	Collagenase Type 1: 0.075%	DMEM	Emre, E, Yuksel, N, Duruksu, G, Pirhan, D, Subasi, C, Erman, G and Karaoz, E:Neuroprotective Effects of Intravitreally Transplanted Adipose Tissue and Bone Marrow-Derived Mesenchymal Stem Cells in an Experimental Ocular Hypertension Model., Cytotherapy 17, 543-59, 2015
小鼠	小鼠, SD, 5 個月	Mesenchymal Stromal Cells	Collagenase Type 2: 0.075%	DMEM	Veronesi, F, Torricelli, P, Della, B, Pagani, S and Fini, M: In Vitro Mutual Interaction Between Tenocytes and Adipose-Derived Mesenchymal Stromal Cells.,Cytotherapy 17, 215-23, 2015
大鼠	大鼠	Adipocytes	Collagenase Type 1: 0.3%	Krebs-Ringer	Ogasawara, J., Izawa, T., Sakurai, T., Shirato, K., Ishibashi, Y., Ohira, Y., Ishida, H., Ohno, H. and Kizaki, T.: Habitual Exercise Training Acts as a Physiological Stimulator for Constant Activation of Lipolytic Enzymes in Rat Primary White Adipocytes., Biochem Biophys Res Commun 464, 348-53, 2015
大鼠	大鼠, 成年的	Stromal vascular	Collagenase Type 1: 0.2%	PBS	Leblanc, A., Nguyen, Q., Touroo, J., Aird, A., Chang, R., Ng, C., Hoying, J. and Williams, S.: Adipose-derived Cell Construct Stabilizes Heart Function and Increases Microvascular Perfusion in an Established Infarct., Stem Cells Transl Med2, 896-905, 2013
大鼠	大鼠, 雄性, 300g	Adipocytes	Collagenase Type 2: 0.33%	RPMI 1640	Thompson, A., Nunez, M., Davidson, R., Horm, T., Schnittker, K., Hart, M., Suarez, A. and Tsao, T.: Mitigation of Isolation-Associated Adipocyte Interleukin-6 Secretion Following Rapid Dissociation of Adipose Tissue., J Lipid Res 53, 2797, 2012
大鼠	大鼠, 路易斯, 雄性	Renal adipose derived cells	Collagenase Type 1: 0.3%	DMEM	Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011
大鼠	大鼠, E18	Neurons	Papain: 0.05%	Neurobasal	Giacomello, M., Girardi, S., Scorzeto, M., Peruffo, A., Maschietto, M., Cozzi, B. and Vassanelli, S.: Stimulation of Ca?+ Signals in Neurons by Electrically Coupled Electrolyte-Oxide-Semiconductor Capacitors., J Neurosci Methods 198, 1, 2011
大鼠	大鼠, SD, 初生	Brown adipocytes	Collagenase Type 4: 0.1%	PBS	Liu, Z., Wang, H., Zhang, Y., Zhou, J., Lin, Q., Wang, Y., Duan, C., Wu, K. and Wang, C.: Efficient Isolation of Cardiac Stem Cells from Brown Adipose., J Biomed Biotechnol 2010, 104296, 2010
			Neutral Protease: 0.1%
			Trypsin: 0.05%
大鼠	大鼠	Adipose Derived Stem	Collagenase Type 1: 0.1%	DMEM	Wei, X., Du, Z., Zhao, L., Feng, D., Wei, G., He, Y., Tan, J., Lee, W., Hampel, H., Dodel, R., Johnstone, B., March, K., Farlow, M. and Du, Y.: IFATS Collection: The Conditioned Media of Adipose Stromal Cells Protect Against Hypoxia-Ischemia-Induced Brain Damage In Neonatal Rats., Stem Cells 27, 478, 2009
大鼠	大鼠	Adipose tissue-derived stem	Collagenase Type 2: 0.075%	DMEM	Tomiyama, K., Murase, N., Stolz, D., Toyokawa, H., O'Donnell, D., Smith, D., Dudas, J., Rubin, J. and Marra, K.: Characterization of Transplanted Green Fluorescent Protein+ Bone Marrow Cells Into Adipose Tissue., Stem Cells 26, 330, 2008
大鼠	大鼠, 大鼠肉瘤, 4 周	Adipocytes	Collagenase: 0.2%	Ham's F12	Aoki, S., Toda, S., Sakemi, T., and Sugihara, H.: Coculture of Endothelial Cells and Mature Adipocytes Actively Promotes Immature Preadipocyte Development In Vitro, Cell Struct Funct 28, 55, 2003
大鼠	大鼠, SD	Adipocytes	Collagenase Type 1: 0.2%	KRHB	Mora, S., Yang, C., Ryder, J., Boeglin, D and Pessin, J: The MEF2A and MEF2D Isoforms are Differentially Regulated in Muscle and Adipose Tissue during States of Insulin Deficiency, Endocrinology 142, 1999, 2001
大鼠	大鼠, SD, 雄性, 4-7周	Brown adipocytes	Deoxyribonuclease I: 0.5%	DMEM	Omatsu-Kanbe, M., and Matsuura, H.: Inhibition of Store-operated Ca2+ Entry by Extracellular ATP in Rat Brown Adipocytes, J Physiol 521 (3), 601, 1999
大鼠	大鼠	Adipocytes	Collagenase Type 2: 0.2%	DMEM /F-12	Serrero, G: Primary Culture in Defined Medium of Adipocyte Precursors,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 11B:6.1, 1995
大鼠	大鼠, SD, 雄性, 130-160 克	Adipose	Collagenase: 0.3%	Kreb's-Ringer bicarbonate buffer modified	Charron, M.J. and Kahn, B.B.: Divergent Molecular Mechanisms for Insulin-Resistant Glucose Transport in Muscle and Adipose Cells In Vivo, J Biol Chem 265, 7994, 1990
大鼠	大鼠, SD, 雄性, 130-160 克	Epididymal fat pads	Collagenase: 0.3%	Kreb's-Ringer bicarbonate buffer modified
大鼠	大鼠, 3 天	Preadipocytes	Collagenase Type 3: 0.10%	Parker Medium 199	Gaben-Cogneville, A., Poussin, B., Chamblier, M., Forgue-Fafitte, M., and Rosselin, G.: Development of Insulin and Epidermal Growth Factor Receptors During the Differentiation of Rat Preadipocytes in Primary Culture, Biochim Biophys Acta 968, 231, 1988
大鼠	大鼠, SD, 不同的重量和年齡	Brown adipocytes	Collagenase: 0.2%	Krebs Ringer bicarbonate buffer	Woodward, Julie A. and Saggerson, E.: Effect of Adenosine Deaminase, N6-Phenylisopropyladenosine and Hypothroidism on the Responsiveness of Rat Brown Adipocytes to Noradrenaline, Biochem J 238, 395, 1986
大鼠	大鼠, SD, 不同的重量和年齡	Interscapular & cervical depots	Soybean Trypsin Inhibitor: 0.3%	Krebs Ringer bicarbonate buffer
大鼠	大鼠, CD, 雄性, 150-200克	Adipocytes	Collagenase: 0.1%	Krebs Ringer bicarbonate buffer	Pessin, J.E., Gitomer, W., Oka, Y., Oppenheimer, C.L., and Czech, M.P.: ?-Adrenergic Regulation of Insulin and Epidermal Growth Factor Receptors in Rat Adipocytes, J Biol Chem 258, 7386, 1983
大鼠	大鼠, CD, 雄性, 150-200克	Epididymal fat pads	Collagenase: 0.1%	Krebs Ringer bicarbonate buffer
大鼠	大鼠,大鼠肉瘤, 白化病, 雄性, 100-140 g	Adipocytes	Collagenase: 0.3%	Kreb's Ringer	Green, A. and Newsholme, E.: Sensitivity of Glucose Uptake and Lipolysis of White Adipocytes of the Rat to Insulin and Effects of Some Metabolites, Biochem J180, 365, 1979
大鼠	大鼠,大鼠肉瘤, 白化病, 雄性, 100-140 g	Epididymal-fat pads	Collagenase: 0.3%	Kreb's Ringer
大鼠	大鼠, 菲舍爾, 344, 雄性, 9 - 13 周	White fat	Collagenase: 0.3%	Kreb's Ringer bicarbonate buffer	Stiles, J.W., Francendese, A.A. and Masoro, E.J.: Influence of Age on Size and Number of Fat Cells in the Epididymal Depot, Am J Physiol 229, 1561, 1975
大鼠	大鼠, 大鼠肉瘤, 雄性, 5 周-16 個月	White fat	Collagenase: 0.05%	Kreb's Ringer bicarbonate buffer	DiGirolamo, M., Mendlinger, S., and Fertig, J.W.: A Simple Method to Determine Fat Cell Size and Number in Four Mammalian Species, Am J Physiol 221, 850, 1971
大鼠	大鼠 (CFE), 白化病, 雌性	Brown fat	Collagenase Type 1: 0.10%	Bicarbonate buffer	Fain, J., Reed, N., and Saperstein, R.: Isolation and Metabolism of Brown Fat Cells, J Biol Chem 242, 1887, 1967
大鼠	大鼠, SD, 雄性, 160-210 克	Fat	Collagenase: 0.3%	Albumin-bicarbonate buffer	Rodbell, M.: Metabolism of Isolated Fat Cells. I. Effects of Hormones on Glucose Metabolism and Lipolysis, J Biol Chem 239 (2), 375, 1964

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