期刊
BIO-PROTOCOL
卷 9, 期 17, 页码 -出版社
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.3351
关键词
Electric cell-substrate Impedance sensing; Wound-healing; Intestinal epithelial cell Monolayers; Ulcerative colitis (UC); Caco2-BBE cells
类别
资金
- NIDDK NIH HHS [R01 DK071594] Funding Source: Medline
- BLRD VA [IK6 BX004476] Funding Source: Medline
Here, we describe an in vitro epithelial wound-healing assay using Electric Cell-Substrate Impedance Sensing (ECIS) technology. The ECIS technology is a real time cell growth assay based on a small (250 mu m diameter) active gold electrode which resistance is measured continuously. When intestinal epithelial cells reach confluency on the gold electrode, resistances reach a plateau. For the wound-healing assays, confluent intestinal epithelial monolayers are subjected to a current of 40 kHz frequency, 1,400 mu A amplitude, and 30-second duration. This kills the cells around the small active gold electrode, causing detachment and generating a wound that is healed by surrounding cells that have not been submitted to the current pulse. Wound healing is then assessed by continuous resistance measurements for approximately 30 h after wound. Both cell wounding and measurements of the subsequent healing process are carried out under computer control that takes online measurements each 30 s and stores the data. ECIS technology can be used to study the underlying causes for impaired mucosal healing and to test the efficacy of drugs in mucosal healing.
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