worthington 软骨组织分离指南

发布者：艾美捷科技发布时间：2022-06-01

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原代细胞分离中，软骨组织分离涉及到软骨细胞、间充质、基质细胞囊泡、关节软骨细胞、骨骺和关节软骨等等的解离通常会使用酶，如何选择合适的酶来完成对应软骨组织的解离，以及原代细胞采集的常规操作， 艾美捷针对软骨组织相关的组织分类解离根据各种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献汇总详细指南：软骨组织分离指南

本技术指南描述了标准实验室程序；为建立细胞分离方案提供了一种合乎逻辑的实验方法；并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南

软骨
种属	种属详情	细胞名称	酶组合推荐，含推荐浓度	培养基	参考文献
牛	Bovine, 18-36 month	软骨细胞 / Chondrocytes	Collagenase Type 1: 0.1%	DMEM	White, R. and Gibson, J.: The Effect of Oxygen Tension on Calcium Homeostasis in Bovine Articular Chondrocytes., J Orthop Surg Res 5, 27, 2010
牛	Bovine, 6-8 week	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.2%	DMEM	Hwang, Y., Sangaj, N. and Varghese, S.: Interconnected Macroporous Poly(ethylene glycol) Cryogels as a Cell Scaffold for Cartilage Tissue Engineering., Tissue Eng Part A 16, 3033-41, 2010
牛	Bovine, 1-3 month	软骨细胞 / Chondrocytes	Collagenase Type 4: 390 u/ml	DMEM	Lima, E., Tan, A., Tai, T., Marra, K., DeFail, A., Ateshian, G. and Hung, C.: Genipin Enhances the Mechanical Properties of Tissue-Engineered Cartilage and Protects Against Inflammatory Degradation When Used as a Medium Supplement., J Biomed Mater Res 91, 692, 2009
牛	Steers, 1-2 years	软骨细胞 / Chondrocytes	Trypsin: 0.20%	HBSS	Mackintosh, D., and Mason, R.: Pharmacological Actions of 17 Beta-oestradiol on Articular Cartilage Chondrocytes and Chondrosarcoma Chondrocytes in the Absence of Oestrogen Receptors, Biochim Biophys Acta 964, 295, 1988
牛	Calf, 1-14 days	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.20%	PBS	Klagsbrun, M.: Large Scale Preparation of Chondrocytes,Methods of Enzymology Vol. 58,, , 560, 1979
犬/狗	Canine	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.3%； Trypsin: 0.25%	Ham's F12	Lee, J., Kim, H., Kim, J., Bae, S., Joo, D., Huh, K., Fang, Y., Jeong, J., Kim, M. and Kim, Y.: Microencapsulation of Pancreatic Islets with Canine Ear Cartilage for Immunoisolation., Transplant Proc 44, 1091-4, 2012
鸡	Chick embryos, HH stage	间充质 / Mesenchymal	Trypsin: 0.1%	DMEM；(see reference)	Wong, M., and Tuan, R.: Nuserum, A Synthetic Serum Replacement, Supports Chondrogenesis of Embryonic Chick Limb Bud Mesenchymal Cells in Micromass Culture, In Vitro Cell Dev Biol 29A, 917, 1993
鸡	Chicken, broiler strain, 8-10 weeks	基质细胞囊泡，骨骺生长板 / Matrix vesicles，Epiphyseal growth plate	Trypsin: 0.1%	Tris-buffered saline	Genge, B., Wu, L. and Wuthier, R.: Differential Fractionation of Matrix Vesicle Proteins: Further Characterization of the Acidic Phospholipid-dependent Ca2+-Binding Proteins, J Biol Chem 265, 4703, 1990
鸡	Chicken, broiler strain, 8-10 week	基质细胞囊泡 / Matrix vesicles	Trypsin: 0.1%	(see reference)	Genge, B.R., Wu, L.N.Y., and Wuthier, R.E.: Identification of Phospholipid-dependent Calcium-binding Proteins as Constituents of Matrix Vesicles, J Biol Chem 264, 10917, 1989
鸡	Chick embryos, White Leghorn, 19 day old	成纤维细胞，上皮样 / Fibroblasts，Epithelial-like	Trypsin: 0.25%	E 199 medium	Gionti, E., Capasso, O., and Cancedda, R.: The Culture of Chick Embryo Chondrocytes and the Control of Their Differentiated Functions in Vitro, J Biol Chem 258 (11), 7190, 1983
鸡	Chick embryos, 19 day old	软骨细胞 / Chondrocytes	Trypsin: 0.75%	Coon's modified F-12	Capasso, O., Gionti, E., Pontarelli, G., Ambesi-Impiobato, F., Nitsch, L., Tajana, G., and Cancedda, R.: The Culture of Chick Embryo Chondrocytes and the Control of Their Differentiated Functions In Vitro, Exp Cell Res 142, 197, 1982
鸡	Chick embryos, White Leghorn	翼芽 / Wing buds	Trypsin: 0.1%	Saline G	Ahrens, P., Solursh, M., and Reiter, R.: Stage-Related Capacity for Limb Chondrogenesis in Cell Culture, Dev Biol 60, 69, 1977
马	Equine	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.15%	DMEM	Visser, J., Levett, P., te Moller, N., Besems, J., Boere, K., van Rijen, M., de Grauw, J., Dhert, W., van Weeren, P. and Malda, J.: Crosslinkable Hydrogels Derived from Cartilage, Meniscus, and Tendon Tissue., Tissue Eng Part A 21, 1195-206, 2015
马	Equine	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.08%	DMEM/F12	Ley, C., Svala, E., Nilton, A., Lindahl, A., Eloranta, M., Ekman, S. and Skioldebrand, E.: Effects of High Mobility Group Box Protein-1, Interleukin-1B, and Interleukin-6 on Cartilage Matrix Metabolism in Three-Dimensional Equine Chondrocyte Cultures., Conn Tissue Res 52, 290-300, 2011
山羊	Goat	软骨细胞 / Chondrocytes	Collagenase Type 2: 2%	DMEM	Bekkers,J, Creemers, L, Tsuchida, A, van Rijen, M, Custers, R, Dhert, W and Saris, D.: One-Stage Focal Cartilage Defect Treatment with Bone Marrow Mononuclear Cells and Chondrocytes Leads to Better Macroscopic Cartilage Regeneration Compared to Microfracture in Goats., Osteoarthritis Cartilage 21, 950-6, 2013
人	Human	软骨细胞 / Chondrocytes	Trypsin: 0.25%； Hyaluronidase: 0.1%； Collagenase Type 2: 1000 u/ml； Collagenase Type 1: 400 u/ml	Ham's F12	Muraglia, A., Nguyen, V., Nardini, M., Mogni, M., Coviello, D., Dozin, B., Strada, P., Baldelli, I., Formica, M., Cancedda, R. and Mastrogiacomo, M.: Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support., Front Bioeng Biotechnol 5, 66, 2017
人	Human	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.15%	DMEM	Andrews, S., Kunze, M., Mulet-Sierra, A., Williams, L., Ansari, K., Osswald, M. and Adesida, A.: Strategies to Mitigate Variability in Engineering Human Nasal Cartilage., Sci Rep 7, 6490, 2017
人	Human	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.15%	DMEM	Capsoni, F., Ongari, A., Lonati, C., Accetta, R., Gatti, S. and Catania, A.: Alpha-Melanocyte-Stimulating-Hormone (a-MSH) Modulates Human Chondrocyte Activation Induced by Proinflammatory Cytokines., BMC Musculoskelet Disord 16, 154, 2015
人	Human	软骨细胞 / Chondrocytes	Collagenase: 0.3%	DMEM	Tamai, M., Nagasao, T., Yanaga, H., Hamamoto, Y., Kogure, T. and Tanaka, Y.: Correction of Secondary Deformity after Nuss Procedure for Pectus Excavatum by Means of Cultured autologous Cartilage Cell Injection., Int J Surg Case Rep 15, 70-3, 2015
人	Human, 52-82 years	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.2%； Pronase: 0.15%	DMEM/F12	Pallu, S., Francin, P., Guillaume, C., Gegout-Pottie, P., Netter, P., Mainard, D., Terlain, B. and Presle, N.: Obesity Affects the Chondrocyte Responsiveness to Leptin in Patients with Osteoarthritis., Arthritis Res Ther 12, R112, 2010
人	Human	滑膜细胞 / Synoviocytes	Collagenase Type 1: 0.4%	DMEM	Kim Wan-Uk, Kwok Seung-Ki, Hong Kyung-Hee, Yoo Seung-Ah, Kong Jin-Sun, Choe Jongseon, Cho Chul-Soo: Soluble Fas ligand inhibits angiogenesis in rheumatoid arthritis, Arthritis Res Ther 9, R42, 2007
人	Human, 26-68 year	半月板和软骨 / Meniscus and cartilage	Collagenase Type 2: 0.15%	DMEM	Marsano A, Millward-Sadler SJ, Salter DM, Adesida A, Hardingham T, Tognana E, Kon E, Chiari-Grisar C, Nehrer S, Jakob M, Martin I: Differential cartilaginous tissue formation by human synovial membrane, fat pad, meniscus cells and articular chondrocytes, Osteoarthritis Cartilage 15, 48-58, 2007
人	Human, 64-83 yr	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.08%	DMEM/F12	Tallheden T, Bengtsson C, Brantsing C, Sjogren-Jansson E, Carlsson L, Peterson L, Brittberg M, and Lindahl A.: Proliferation and Differentiation Potential of Chondrocytes from Osteoarthritic Patients, Arthritis Res Ther 7(3), R560, 2005
人	Human, adult	滑膜 / Synovial	Collagenase: 0.15%； Hyaluronidase: 0.1%； Deoxyribonuclease I: 0.015%	DMEM	Liagre B, Vergne-Salle P, Corbiere C, Charissoux JL, and Beneytout JL.: Diosgenin, a Plant Steroid, Induces Apoptosis in Human Rheumatoid Arthritis Synoviocytes with Cyclooxygenase-2 Overexpression, Arthritis Res Ther 6(4), R373, 2004
人	Human, adult	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.15%	DMEM	Jakob M, Demarteau O, Schafer D, Stumm M, Heberer M, and Martin I.: Enzymatic Digestion of Adult Human Articular Cartilage Yields a Small Fraction of the Total Available Cells, Conn Tissue Res 44, 173, 2003
人	Human, adults	滑膜细胞 / Synoviocytes	Collagenase Type 1: 0.1%	RPMI	McEvoy A., Murphy E., Ponnio T., Conneely O., Bresnihan B., FitzGerald O., and Murphy E.: Activation of Nuclear Orphan Receptor NURR1 Transcription by NF-kappa B and Cyclic Adenosine 5'-Monophosphate Response Element-binding Protein in Rheumatoid Arthritis Synovial Tissue, J Immunol 168(6), 2979, 2002
人	Human, 15-60 years	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.2%	DMEM	Rotter N, Bonassar LJ, Tobias G, Lebl M, Roy AK, Vacanti CA: Age dependence of cellular properties of human septal cartilage: implications for tissue engineering, Arch Otolaryngol Head Neck Surg 127, 1248-52, 2001
人	Human	滑膜成纤维细胞 / Synovial fibroblasts	Collagenase: 0.4%	DMEM	Sarkissian M, Lafyatis R: Integrin engagement regulates proliferation and collagenase expression of rheumatoid synovial fibroblasts, J Immunol 162, 1772-9, 1999
人	Human	隔膜软骨细胞 / Septal chondrocytes	Collagenase Type 2: 0.2%； Hyaluronidase: 0.01%； Deoxyribonuclease I: 0.015%	DMEM/F12	Dunham B P, Koch R J: Basic fibroblast growth factor and insulinlike growth factor I support the growth of human septal chondrocytes in a serum-free environment, Arch Otolaryngol Head Neck Surg 124, 1325-30, 1998
人	Human w/ rheumatoid arthritis	滑膜组织 / Synovial tissue	Trypsin: 0.05%	DMEM (see reference)	Dayer, J., Krane, S., Russell, R., and Robinson, D.: Production of Collagenase and Prostaglandins by Isolated Adherent Rheumatoid Synovial Cells, Proc Natl Acad Sci U S A 73 (3), 945, 1976
人	Human, 13-62 years	关节软骨细胞 / Articular chondrocytes	Trypsin: 0.2%	BSS	Srivastava, V.M.L., Malemud, C.J., Hough, A.J., Bland, J.H., and Sokoloff, L.: Preliminary Experience with Cell Culture of Human Articular Chondrocytes, Arthritis Rheum 17, 165, 1974
人	Human, 26-84 years	软骨细胞 / Chondrocytes	Collagenase:	GBSS	Manning, W.K., and Bonner, W.M.: Isolation and Culture of Chondrocytes From Human Adult Articular Cartilage, Arthritis Rheum 10, 235, 1967
小鼠	Mouse, 2-5 day	软骨细胞 / Chondrocytes	Trypsin: 0.25%； Collagenase Type 1: 86.5 u/ml	DMEM	Otsuru, S., Hofmann, T., Raman, P., Olson, T., Guess, A., Dominici, M. and Horwitz, E.: Genomic and Functional Comparison of Mesenchymal Stromal Cells Prepared Using Two Isolation Methods., Cytotherapy 17, 262-70, 2015