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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。
脂肪組織消化手冊(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 | 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 |
| (還有小鼠, 倉鼠, 兔, 羔羊, 豚鼠) | |||||
| 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 |
| (還有鼠, 倉鼠, 沙鼠, 兔, 羔羊) | |||||
| 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 |
| (還有鼠, 沙鼠, 兔, 羔羊, 豚鼠) | |||||
| 倉鼠 | 倉鼠, 5 周-12 個(gè)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 |
| Deoxyribonuclease I: 0.002% | |||||
| 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 |
| Neutral Protease: | |||||
| 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(gè)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(jì) | 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(gè)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 |
| Epididymal fat pads | |||||
| 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 |
| Interscapular & cervical depots | Soybean Trypsin Inhibitor: 0.3% | ||||
| 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 |
| Epididymal fat pads | |||||
| 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 |
| Epididymal-fat pads | |||||
| 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(gè)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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