worthington 軟骨組織分離指南

原代細胞分離中,軟骨組織分離涉及到軟骨細胞、間充質、基質細胞囊泡、關節軟骨細胞、骨骺和關節軟骨等等的解離通常會使用酶,如何選擇合適的酶來完成對應軟骨組織的解離,以及原代細胞采集的常規操作, 艾美捷針對軟骨組織相關的組織分類解離根據各種屬、不同的細胞、酶組合推薦,推薦濃度、培養基以及參考文獻匯總詳細指南:

腦組織分離指南

 

本技術指南描述了標準實驗室程序; 為建立細胞分離方案提供了一種合乎邏輯的實驗方法; 并列出了許多組織特異性參考資料,以幫助開發有效的方法。更多其他組織的解離指南,點擊查看:各組織分離指南

軟骨
種屬 種屬詳情 細胞名稱 酶組合推薦,含推薦濃度 培養基 參考文獻
Bovine, 18-36 month 軟骨細胞 / 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
Bovine, 6-8 week 軟骨細胞 / 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
Bovine, 1-3 month 軟骨細胞 / 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
Steers, 1-2 years 軟骨細胞 / 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
Calf, 1-14 days 軟骨細胞 / Chondrocytes Collagenase Type 2: 0.20% PBS Klagsbrun, M.: Large Scale Preparation of Chondrocytes,Methods of Enzymology Vol. 58,, , 560, 1979
犬/狗 Canine 軟骨細胞 / 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
Chick embryos, HH stage 間充質 / 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
Chicken, broiler strain, 8-10 weeks 基質細胞囊泡,骨骺生長板 / 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
Chicken, broiler strain, 8-10 week 基質細胞囊泡 / 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
Chick embryos, White Leghorn, 19 day old 成纖維細胞,上皮樣 / 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
Chick embryos, 19 day old 軟骨細胞 / 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
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
Equine 軟骨細胞 / 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
Equine 軟骨細胞 / 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 軟骨細胞 / 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
Human 軟骨細胞 / 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
Human 軟骨細胞 / 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
Human 軟骨細胞 / 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
Human 軟骨細胞 / 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
Human, 52-82 years 軟骨細胞 / 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
Human 滑膜細胞 / 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
Human, 26-68 year 半月板和軟骨 / 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
Human, 64-83 yr 軟骨細胞 / 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
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
Human, adult 軟骨細胞 / 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
Human, adults 滑膜細胞 / 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
Human, 15-60 years 軟骨細胞 / 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
Human 滑膜成纖維細胞 / 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
Human 隔膜軟骨細胞 / 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
Human w/ rheumatoid arthritis 滑膜組織 / 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
Human, 13-62 years 關節軟骨細胞 / 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
Human, 26-84 years 軟骨細胞 / 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 軟骨細胞 / 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
小鼠 Mouse, 1 day 軟骨細胞 / Chondrocytes Collagenase: 0.2% DMEM Terpstra, L, Prud'homme, J, Arabian, A, Takeda, S, Karsenty, G, Dedhar, S, and St-Arnaud, R.: Reduced Chondrocyte Proliferation and Chondrodysplasia in Mice Lacking the Integrin-linked Kinase in Chondrocytes, J Cell Biol 162, 139, 2003
綿羊 Sheep, 2 month 軟骨細胞 / Chondrocytes Collagenase Type 2: 0.3% Ham's F-12 Kojima Koji, Bonassar Lawrence J, Roy Amit K, Mizuno Hirokazu, Cortiella Joaquin, Vacanti Charles A: A Composite Tissue-Engineered Trachea Using Sheep Nasal Chondrocyte and Epithelial Cells, FASEB J 17, 823-8, 2003
Porcine, 1 year 軟骨細胞 / Chondrocytes Collagenase Type 1: 0.2% DEMEM Chowdhury, T., Schulz, R., Rai, S., Thuemmler, C., Wuestneck, N., Bader, A and Homandberg, G: Biomechanical Modulation of Collagen Fragment-Induced Anabolic and Catabolic Activities in Chondrocyte/Agarose Constructs., Arthritis Res Ther 12, R82, 2010
Porcine, 2-4 month 軟骨 / Chondrons Neutral Protease: 0.3%;
Collagenase: 0.2%
PBS Graff RD, Lazarowski ER, Banes AJ, Lee GM: ATP release by mechanically loaded porcine chondrons in pellet culture, Arthritis Rheum 43, 1571-9, 2000
Rabbit, New Zealand, 1.2-1.4 kg 軟骨細胞 / Chondrocytes Collagenase Type 2: 0.025%;
Pronase: 0.2%
DMEM Ju, X., Deng, M., Ao, Y., Yu, C., Wang, J., Yu, J., Cui, G. and Hu, Y.: Protective Effect of Sinomenine on Cartilage Degradation and Chondrocytes Apoptosis., Yakugaku Zasshi 130, 1053-60, 2010
Rabbit, New Zealand, 1.8-2.3kg 軟骨細胞 / Chondrocytes Hyaluronidase: .05%;
Collagenase Type 2: 0.2%;
Trypsin: 0.2%
Gey's solution Mehraban F, Tindal MH, Proffitt MM, Moskowitz RW.: Temporal pattern of cysteine endopeptidase (cathepsin B) expression in cartilage and synovium from rabbit knees with experimental osteoarthritis: gene expression in chondrocytes in response to interleukin-1 and matrix depletion, Ann Rheum Dis 56, 108, 1997
Rabbit, New Zealand, white, 4-6 wk & 22-25 wk 軟骨細胞 / Chondrocytes Protease XIV: 5 mg/g of tissue Ham's F-12 Plaas, A., Sandy, J., and Kimura, J.: Biosynthesis of Cartilage Proteoglycan & Link Protein Articular Chondrocytes, J Biol Chem 263, 7560, 1988
Rabbit, white, male, 8 weeks 軟骨細胞 / Chondrocytes Trypsin: 0.2% Gey's BSS Benya, P.D., Padilla, S.R., and Nimni, M.E.: The Progeny of Rabbit Articular Chondrocytes Synthesize Collagen Types I and III and Type I Trimer, but Not Type II, Biochemistry 16, 865, 1977
Rabbit, New Zealand white or Dutch, 1 week (also human, newborn) 軟骨細胞 / Chondrocytes Trypsin: 0.1% Saline G, CMF Schindler, F.H., Ose, M.A., and Solursh, M.: The Synthesis of Cartilage Collagen by Rabbit and Human Chondrocytes in Primary Cell Culture, In Vitro 12, 44, 1976
Rabbit, New Zealand white, immature, 2.25 - 3.3 Kg 關節軟骨細胞,透明 / Articular chondrocytes,Hyaline Trypsin: 0.2% Gey's BSS Green, J.R., and William, T.: Articular Cartilage Repair. Behavior of Rabbit Chondrocytes During Tissue Culture and Subsequent Allografting, Clin Orthop Relat Res , 237, 1976
Rabbit, New Zealand white, male, 250-350 g 骨骺,關節軟骨 / Epiphyseal,Articular cartilage Trypsin: 0.25% Eagle's basal medium Bentley, G., and Greer , R.: Homotransplantation of Isolated Epiphyseal and Articular Cartilage Chondrocytes into Joint Surfaces of Rabbits, Nature 230, 385, 1971
大鼠 Rat 軟骨細胞 / Chondrocytes Collagenase Type 2: 0.15%;
Trypsin: 0.25%
DMEM/F12 Mei, L., Shen, B., Ling, P., Liu, S., Xue, J., Liu, F., Shao, H., Chen, J., Ma, A. and Liu, X.: Culture-expanded Allogenic Adipose Tissue-derived Stem Cells Attenuate Cartilage Degeneration in an Experimental Rat Osteoarthritis Model., PLoS ONE 12, e0176107, 2017
大鼠 Rat, SD, young, 100-120 g 軟骨細胞 / Chondrocytes Trypsin: 0.2% Ham's F-12 medium Shimomura, Y., Yoneda, T., and Suzuki, F.: Osteogenesis by Chondrocytes from Growth Cartilage of Rat Rib, Calcif Tissue Res 19, 179, 1975

 

艾美捷科技優勢代理品牌

發表評論

:?: :razz: :sad: :evil: :!: :smile: :oops: :grin: :eek: :shock: :???: :cool: :lol: :mad: :twisted: :roll: :wink: :idea: :arrow: :neutral: :cry: :mrgreen: