Rajan Navneeta, Habermehl Jason, Coté Marie-France, Doillon Charles J, Mantovani Diego
Laboratory for Biomaterials and Bioengineering, Laval University and University Hospital Research Center, Quebec City, Quebec G1K 7P4, Canada.
Nat Protoc. 2006;1(6):2753-8. doi: 10.1038/nprot.2006.430.
Collagen is a widely investigated extracellular matrix material with extensive potentials in the field of tissue engineering. This protocol describes a method to prepare reconstituted collagen that can be ready-to-use, storable and suitable for further in vitro and in vivo investigations. Type I collagen was extracted from rat tail tendons and processed in acetic acid solution to obtain sterile soluble collagen. At first, crude collagen was dissolved in acetic acid, then frozen at -20 degrees C and lyophilized to obtain a sponge, which could be stored at -80 degrees C. Lyophilized collagen was then dispersed in acetic acid to obtain a sterile solution of collagen at targeted concentrations. The whole low-cost process from the extraction to the final sterile solution takes around 2-3 weeks. The collagen solution, once neutralized, has the potential to be used to produce gels or scaffolds, to deposit thin films on supports and to develop drug delivery systems.
胶原蛋白是一种在组织工程领域具有广泛潜力且被广泛研究的细胞外基质材料。本方案描述了一种制备可直接使用、可储存且适合进一步体外和体内研究的重组胶原蛋白的方法。从大鼠尾腱中提取I型胶原蛋白,并在乙酸溶液中进行处理以获得无菌可溶性胶原蛋白。首先,将粗制胶原蛋白溶解在乙酸中,然后在-20℃下冷冻并冻干以获得海绵体,该海绵体可储存在-80℃。然后将冻干的胶原蛋白分散在乙酸中以获得目标浓度的无菌胶原蛋白溶液。从提取到最终无菌溶液的整个低成本过程大约需要2至3周。一旦中和,胶原蛋白溶液就有潜力用于制备凝胶或支架、在载体上沉积薄膜以及开发药物递送系统。