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脂肪組織消化手冊

更新時間:2016-02-17點擊次數(shù):1571

世界*實驗材料供應商 Worthington Biochemical 正式授權上海起發(fā)為其中國代理, Worthington Biochemical 在一直是行業(yè)的*,一直為廣大科研客戶提供zui為的產(chǎn)品和服務,上海起發(fā)一直秉承為中國科研客戶帶來的產(chǎn)品,的服務,簽約 Worthington Biochemical 就是為了給廣大科研客戶帶來更加完善的產(chǎn)品和服務,您的滿意將是我們zui大的收獲

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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(jīng)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(jīng)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(jù)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(yǎng)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(jīng)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(shù)據(jù)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(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, 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(nèi)皮組織

上皮組織

心臟

淋巴結

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(yǎng)基參考文獻
AdipocytesCollagenase Type 1: 40 u/mlKrebs-Ringer bicarbonateYang, Y., Baldwin, R.: Preparation and Metabolism of Isolated Cells from Bovine Adipose Tissue, J Dairy Sci 56, 350, 1973
Renal adipose derived cellsCollagenase Type 1: 0.3%DMEMBasu, 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, 20-25公斤Adipose stem cellsCollagenase: see referenceMedia-199Fischer, 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
White fatCollagenase: 0.05%Kreb's Ringer bicarbonate bufferDiGirolamo, 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
Adipose derived stem cellsCollagenase Type 1: 0.1%PBSVidal, 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, 1-5 年Adipose derived stem cellsCollagenase Type 1: 0.1%PBSVidal, 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, 大西洋鮭魚PreadipocytesCollagenase Type 1: 0.1%HBSSTodorcevic, 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 fatCollagenase Type 1: 0.10%Bicarbonate bufferNedergaard, 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 fatCollagenase Type 1: 0.10%Bicarbonate bufferNedergaard, 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 fatCollagenase Type 1: 0.10%Bicarbonate bufferNedergaard, 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 個月White fatCollagenase: 0.05%Kreb's Ringer bicarbonate bufferDiGirolamo, 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
Mesenchymal Stromal CellsCollagenase Type 4: 0.2%PBSChoudhery,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
Adipose derived stem cellsCollagenase Type 2: 0.1%HBSSSatish, 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, 女性Adipose StromalCollagenase Type 1: 0.075%DMEMKinoshita, 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, 成年的Adipose derived mesenchymal stemCollagenase Type 2: 0.1%see referenceAl-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, 成年的Adipose derived stem cellsCollagenase Type 1: 0.15%DMEMKoellensperger, 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
Mesenchymal Stromal CellsCollagenase Type 1: 0.1%PBSNajar, 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
Adipose stromal stemCollagenase animal free: 200 u/mlDMEM/Hams F-12Carvalho, 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
Stromal Vascular FractionCollagenase Type 1: 0.075%PBSDoi, 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
Adipose-derived stem cellsCollagenase Type 1: 0.1%PBSCervelli, 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
PerivascularCollagenase Type 2: 0.1%DMEMJames, 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
Adipose derived stromal vascularCollagenase Type 1: 0.1%PBSGentile, 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
Stromal vascular, adipocytesCollagenase Type 1: 0.1%DMEMGentile, 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
Chorionic mesenchymalCollagenase Type 1: 0.1%MEMNazarov, 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, 成年的StromovascularCollagenase Type 1: 0.1%PBS with BSAHagman, 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
Adipose derived stromalCollagenase Type 1: 0.1%PBSNaaijkens, 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
Adipose derived stemCollagenase Type 1: 0.1%DMEM/F-12Wu, 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, 男性 40-60歲Adipose derived stem cellsCollagenase: 0.25%PBSBlasi, 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%
Renal adipose derived cellsCollagenase Type 1: 0.3%DMEMBasu, 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
Adipose derived mesenchymalCollagenase Type 1: 0.1%L-DMEMYang, 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
Adipose derived stemCollagenase Type 1: 0.1%DMEM/Ham's F-12Yu, 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
AdipocytesCollagenase Type 1: 0.1%RPMIBasu, 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
AdipocytesCollagenase Type 1: 0.1%KRBPeters, 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
Adipose tissue-derived stemCollagenase Type 2: 1.0%DMEM/F12Tan, 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
Stromal vascularCollagenase Type 2: 0.1%HBSSZimmerlin, 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
Adipose stromalCollagenase Type 1: 0.1%DMEM/Hams F-12Yu, 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
AdipocytesCollagenase: 0.075%DMEMSuga, 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
Adipose Stromal CellCollagenase Type 1: 0.1%DMEMCai, 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
Adipose tissue-derived stemCollagenase Type 2: 1.0%DMEM/F12Tan, 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
Adipose derived stem cellsCollagenase Type 1: 0.1%DMEMTandon, 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
Adipose derived stromal cellsCollagenase Type 1: see referenceDMEMTraktuev, 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
StromalCollagenase Type 1: 0.2%PBSMinana, 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
StromalCollagenase Type 1: 0.1%DMEMNie, 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: 
Stem CellsCollagenase: 280 u/mlD-PBSPilgaard, 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
AdipocytesCollagenase Type 1: 0.2%HBSSBujalska 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
Adipose derived stromal cellsCollagenase Type 1: 0.15%DMEMSchaffler 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
Adipose derived stem cellsCollagenase Type 1: 0.1%DMEM/F12Kilroy, 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
Mesenchymal stemCollagenase Type 1: 0.1%HBSSJeon 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
PreadipocytesCollagenase Type 1: 196 u/mlM199Koellensperger, 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
Adipocytes, stromal vascularCollagenase: 0.2%HBSSBoquest 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
Multipotent adipose derived stemCollagenase: 0.2%DMEMRodriguez 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
成年的 stem cellsCollagenase Type 1: 0.1%PBSDevireddy, 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, 成年的 , 肥胖的AdipocytesCollagenase Type 2: 0.1%DMEM/F12Seboek, 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, 成年的, 女性PreadipocytesCollagenase Type 1: 0.2%DMEM/F-12Quickler, 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,成年的AdipocytesCollagenase Type 1: 0.13%see referenceFain 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
Stromal vascular, adipocytesCollagenase: 0.2%DMEM/F12Planat-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
Stromal vascular, adipocytes, stemCollagenase: 300 u/mlPBSMiranville 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, 成年的, 女性, 非糖尿病的AdipocytesCollagenase Type 2: 0.05%DMEMGesta, 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
AdipocytesCollagenase Type 1: see referenceSalinePatwardhan, 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, 成年的, 男性和女性AdipocytesCollagenase Type 1: 0.2%DMEM/F12McTernan, 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
Processed lipoaspi鼠e cellsCollagenase Type 1: 0.075%PBSZuk 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
AdipocytesCollagenase Type 1: 0.2%HBSSMcTernan 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, 成年的, 男性和女性AdipocytesCollagenase Type 1: 0.1%DMEM/Ham's F-12Gottschling-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, 成年的, 男性和女性AdipocytesCollagenase Type 1: 0.1%DMEM/F12Zhang, 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, 非糖尿病的, 男性AdipocytesCollagenase Type 1: 0.05%Kreb's Ringer bicarbonate bufferAnderson, O., Gliemann, J., and Gammeltoft: Receptor Binding and Biological Effect of Insulin in Human Adipocytes, Diabetologia 13, 589, 1977
小鼠小鼠AdipocytesCollagenase: 0.1%KrebsJang, 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 VascularCollagenase Type 2: 0.2%RPMI-1640Kondo, 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 stemCollagenase Type 2: 0.1%DMEMTakahashi, 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 progenitorCollagenase Type 2: 0.2%HBSSHan, 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AdipocytesCollagenase Type 1: 0.2%HBSSDe 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 adipocytesCollagenase Type 2: 0.1%DMEMWong, 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 CellCollagenase Type 2: 0.075%DMEMThangarajah, 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 stemCollagenase Type 1: 0.075%Modified EaglesCho, 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/6JAdipocytesCollagenase Type 1: 0.15%DMEM/F12Aoyagi 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 cellsCollagenase Type 2: 0.075%PBSCowan 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, adipocytesCollagenase: 0.2%DMEM/F12Planat-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AdipocytesCollagenase: see referenceDMEMRuan, 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AdipocytesCollagenase: 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 endothelialCollagenase: 0.2%PBSLaunder, 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, 雌性, 一年以下Adipose mesenchymal stemCollagenase Type 1: 0.1%DMEMWilliams, 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, 雜交繁育, 雄性 1-4 天AdipocytesCollagenase Type 1: 0.2%DMEM/F12Ramsay, T.G.: Porcine Leptin Inhibits Lipogenesis in Porcine Adipocytes, J Anim Sci 81, 3008, 2003
豬, 雜交繁育的AdipocytesCollagenase Type 1: 0.067%Krebs-RingerLiang, W. and Mills, S.: Quantitative Analysis of beta-Adrenergic Receptor Subtypes in Pig Tissues., J Anim Sci 80, 963-70, 2002
豬, 8-9 周AdiposeCollagenase Type 1: 300 u/mlHEPESDing, 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, 初生AdipocytesCollagenase Type 1: 0.3%Krebs-Ringer bicarbonate albuminWang Y, Fried SK, Petersen RN, Schoknecht PA: Somatotropin regulates adipose tissue metabolism in neonatal swine, J Nutr 129, 139-45, 1999
豬, 雜交繁育, 1-3 天Adipose, Stromal-vascularCollagenase Type 1: 0.2%DMEM/F12Suryawan, 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 CellsCollagenase Type 1: 0.075%DMEMEmre, 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 CellsCollagenase Type 2: 0.075%DMEMVeronesi, 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AdipocytesCollagenase Type 1: 0.3%Krebs-RingerOgasawara, 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 vascularCollagenase Type 1: 0.2%PBSLeblanc, 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, 300gAdipocytesCollagenase Type 2: 0.33%RPMI 1640Thompson, 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 cellsCollagenase Type 1: 0.3%DMEMBasu, 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, E18NeuronsPapain: 0.05%NeurobasalGiacomello, 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 adipocytesCollagenase Type 4: 0.1%PBSLiu, 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 StemCollagenase Type 1: 0.1%DMEMWei, 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 stemCollagenase Type 2: 0.075%DMEMTomiyama, 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AdipocytesCollagenase: 0.2%Ham's F12Aoki, 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, SDAdipocytesCollagenase Type 1: 0.2%KRHBMora, 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 adipocytesDeoxyribonuclease I: 0.5%DMEMOmatsu-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AdipocytesCollagenase Type 2: 0.2%DMEM /F-12Serrero, 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AdiposeCollagenase: 0.3%Kreb's-Ringer bicarbonate buffer modifiedCharron, 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PreadipocytesCollagenase Type 3: 0.10%Parker Medium 199Gaben-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 adipocytesCollagenase: 0.2%Krebs Ringer bicarbonate bufferWoodward, 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 depotsSoybean Trypsin Inhibitor: 0.3%
大鼠大鼠, CD, 雄性, 150-200克AdipocytesCollagenase: 0.1%Krebs Ringer bicarbonate bufferPessin, 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 gAdipocytesCollagenase: 0.3%Kreb's RingerGreen, 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 fatCollagenase: 0.3%Kreb's Ringer bicarbonate bufferStiles, 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 fatCollagenase: 0.05%Kreb's Ringer bicarbonate bufferDiGirolamo, 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 fatCollagenase Type 1: 0.10%Bicarbonate bufferFain, 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FatCollagenase: 0.3%Albumin-bicarbonate bufferRodbell, 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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