Yue Patrick Y K, Leung Emily P Y, Mak N K, Wong Ricky N S
1Department of Biology, Faculty of Science, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR.
J Biomol Screen. 2010 Apr;15(4):427-33. doi: 10.1177/1087057110361772. Epub 2010 Mar 5.
Cell migration plays a key role in both normal physiological and pathological conditions. The study of cell migration and its underlying mechanisms is of great significance in various fields of research, including basic biology and pharmaceutical development. The cell migration or scratch wounding assay is an easy and economical in vitro method that allows researchers to assess a large number of testing compounds. Even though this simple assay has been used for decades, researchers are still trying to modify such experimental protocols and wounding devices. In this study, an 8-channel mechanical "wounder" was designed for performing a cell migration assay, particularly in a 96-well culture plate format. With special designs of a guiding bar and adjustable pins for use with disposable pipette tips, this wounder confined the scratch area within the center of each well to ensure a perfect contact between the pins and the well surface. As a result, this mechanical wounder produces a uniform denudation of a cell monolayer in a 96-well plate with a wound size of around 600 microm. Using this improved wounding device, the effects of epidermal growth factor and DL-alpha-difluoromethylornithine on the reepithelialization of rat intestinal epithelial cells (IEC-6) and serum on the wound recovery of human umbilical vein endothelial cells were demonstrated. This wounder facilitates cell migration study and can be applicable for multiple sample analysis.
细胞迁移在正常生理和病理状况中均发挥着关键作用。细胞迁移及其潜在机制的研究在包括基础生物学和药物研发在内的各个研究领域都具有重大意义。细胞迁移或划痕损伤试验是一种简便且经济的体外方法，可让研究人员评估大量受试化合物。尽管这种简单的试验已使用了数十年，但研究人员仍在尝试改进此类实验方案和损伤装置。在本研究中，设计了一种8通道机械“损伤器”用于进行细胞迁移试验，特别是以96孔培养板形式。通过导向杆和与一次性移液器吸头配合使用的可调节销的特殊设计，该损伤器将划痕区域限制在每个孔的中心，以确保销与孔表面完美接触。结果，这种机械损伤器在96孔板中产生均匀的细胞单层剥脱，划痕大小约为600微米。使用这种改进的损伤装置，证明了表皮生长因子和DL-α-二氟甲基鸟氨酸对大鼠肠上皮细胞（IEC-6）再上皮化的影响以及血清对人脐静脉内皮细胞伤口愈合的影响。这种损伤器便于细胞迁移研究，可适用于多个样本分析。
