Cao Zhiyi, Saravanan Chandrassegar, Chen Wei-Sheng, Panjwani Noorjahan
New England Eye Center and Department of Ophthalmology, Tufts University, 136 Harrison Avenue, Boston, MA, 02111, USA.
Methods Mol Biol. 2015;1207:317-26. doi: 10.1007/978-1-4939-1396-1_21.
Re-epithelialization is a crucial step for wound healing. As galectins play important roles in re-epithelialization, we describe here protocols for in vivo, ex vivo and in vitro examination of the role of galectins in cell migration and in re-epithelialization of wounds. For in vivo models, mouse corneas are wounded by a variety of techniques and the rate of re-epithelialization is quantified. For ex vivo organ culture models, mouse corneas are wounded in situ, the eyes are enucleated, the eyeballs are cultured in the presence or absence of galectins and the rate of re-epithelialization is quantified. For cell cultured-based in vitro assays, we examine formation of lamellipodia and activation of focal adhesion kinase in various epithelial cells.
再上皮化是伤口愈合的关键步骤。由于半乳糖凝集素在再上皮化过程中发挥重要作用，我们在此描述体内、体外和离体检测半乳糖凝集素在细胞迁移和伤口再上皮化中作用的实验方案。对于体内模型，通过多种技术损伤小鼠角膜，并对再上皮化速率进行定量分析。对于离体器官培养模型，原位损伤小鼠角膜，摘除眼球，在有或无半乳糖凝集素的情况下培养眼球，并对再上皮化速率进行定量分析。对于基于细胞培养的体外分析，我们检测各种上皮细胞中片状伪足的形成和粘着斑激酶的激活情况。
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