Poon Christopher J, Tan Shaun S, Boodhun Sholeh W, Abberton Keren M, Morrison Wayne A
O'Brien Institute Department of SVI, St. Vincent's Institute of Medical Research, Melbourne, Australia.
Bio Protoc. 2017 Feb 5;7(3):e2128. doi: 10.21769/BioProtoc.2128.
Hydrogels are an ideal medium for the expansion of cells in three dimensions. The ability to induce cell expansion and differentiation in a controlled manner is a key goal in tissue engineering. Here we describe a detailed method for producing hydrogels from soft tissues with an emphasis on adipose tissue. In this method, soluble, extractable proteins are recovered from the tissue and stored while the remaining insoluble tissue is processed and solubilised. Once the tissue has been sufficiently solubilised, the extracted proteins are added. The resulting product is a thermosensitive hydrogel with proteins representative of the native tissue. This method addresses common issues encountered when working with some biomaterials, such as high lipid content, DNA contamination, and finding an appropriate sterilisation method. Although the focus of this article is on adipose tissue, using this method we have produced hydrogels from other soft tissues including muscle, liver, and cardiac tissue.
水凝胶是细胞进行三维扩增的理想介质。以可控方式诱导细胞扩增和分化的能力是组织工程的一个关键目标。在此,我们描述一种从软组织制备水凝胶的详细方法,重点是脂肪组织。在该方法中,从组织中回收可溶性、可提取的蛋白质并储存起来,同时对剩余的不溶性组织进行处理并使其溶解。一旦组织充分溶解,就加入提取的蛋白质。所得产物是一种具有代表天然组织蛋白质的热敏水凝胶。该方法解决了使用某些生物材料时遇到的常见问题,如高脂含量、DNA污染以及寻找合适的灭菌方法。尽管本文重点是脂肪组织,但使用此方法我们已从包括肌肉、肝脏和心脏组织在内的其他软组织制备出了水凝胶。