产品名称 | 可溶性I型胶原 |
应用 | 用于传统的二维(2D)系统以及三维(3D)凝胶中的支架,用于模拟成纤维细胞和软骨细胞中的细胞生长。 |
规格 | 4 mg/ml x 12.5 ml (sterile filtered) |
形态 | 溶于0.01M HCL |
来源 | 猪 |
内毒素 | <1 EU/ml |
纯度 | >95% I 胶原 |
保存 | 4℃ |
稳定性 | 6月 |
操作流程 | 包被(注:具体的包被的条件需要根据培养系统优化) 1.用0.02mol/L的盐酸稀释4mg/ml的胶原到50-100ug/ml。 2.温和混匀稀释后的溶液。 3.取适量的稀释后的胶原加入孔内或板内。 注:保证整个表面都是包被的。 4. 37℃或室温孵育1-2小时 5. 弃掉所有溶液。 6.用培养基或者PBS漂洗包被表面。 7.包被好的孔或板2-8℃保存或者空气中晾干(如果保持无菌) 三维凝胶制备流程:
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参考文献 | 1. M. Shoulders, R. Raines, Collagen structure and stability. Annu Rev Biochem 78, 929-58 (2009). 2. W. Friess, Collagen--biomaterial for drug delivery. Eur J Pharm Biopharm 45, 113-36 (1998). 3. Taubenberger, M. Woodruff, H. Bai, D. Muller, D. Hutmacher, The effect of unlocking RGD-motifs in collagen I on pre-osteoblast adhesion and differentiation. Biomaterials 31, 2827-35 (2010). 4. D. Gullberg, K. Gehlsen, D. Turner, K. Ahlén, L. Zijenah, et al., Analysis of alpha 1 beta 1, alpha 2 beta 1 and alpha 3 beta 1 integrins in cell--collagen interactions: identification of conformation dependent alpha 1 beta 1 binding sites in collagen type I. EMBO J 11, 3865-73 (1992). 5. J. Bard, E. Hay, The behavior of fibroblasts from the developing avian cornea. Morphology and movement in situ and in vitro. J Cell Biol 67, 400-18 (1975). 6. H. Kleinman, R. Klebe, G. Martin, Role of collagenous matrices in the adhesion and growth of cells. The Journal of cell biology 88, 473-485 (1981). 7. J. Tomasek, E. Hay, Analysis of the role of microfilaments and microtubules in acquisition of bipolarity and elongation of fibroblasts in hydrated collagen gels. J Cell Biol 99, 536-49 (1984). 8. D. Karamichos, N. Lakshman. W.Petroll, Regulation of corneal fibroblast morphology and collagen reorganization by extracellular matrix mechanical properties. Investigative ophthalmology & visual science 48, 5030-5037 (2007). 9. S. Oliveira, R. Ringshia, R. Legeros, E. Clark, M. Yost, et al., An improved collagen scaffold for skeletal regeneration. J Biomed Mater Res A 94, 371-9 (2010). |
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