Project Details
Abstract
Both collective cell migration, referred as leading cells drive other less active cells to
move toward one direction, and physical interactions between the cell- cell and cell-
substratum play important roles in the wound-healing process. Although the negative-pressure
wound therapy successfully treat difficult chronic wounds, the exact mechanisms are not fully
understood. Electric cell-substrate impedance sensing (ECIS) is a technique to continuously
monitor electric activities of cell migration on the exposed electrode. The optical phase and
wide visual field microscope tracks spatial correlations between embedded fluorescent
particles and cells to estimate the cell-substratum traction force. The purpose of this study
was to use ECIS and traction force microscopy to quantify changes of the cell-cell
adhesion, cell membrane morphologies, and cell-substratum traction force at the
pressure of 635 mmHg. Relations between the above changes and the
epithelial-mesenchymal transition in human keratinocytes (HaCaT cell line) were also
investigated.
First year: The electroporation will be given to wound the monolayer cells in the ECIS
electrode array. The wound-healing process at the ambient pressure and the pressure of 635
mmHg will be continuously recorded in the frequency range of 22.5- 64 kHz from five hours
before wounding to 12 hours after wound closure. The intercellular resistance (Rb), the
cell-substratum resistance (α), and the membrane capacitance (Cm) will be studied during the
wound-healing process.
Second year: To observe the live cell images at different pressures, we planned to put the
Lumascope 500 (Etaluma Inc, Carlsbad, CA, USA) into the negative-pressure incubating
chamber to build the real-time live cell fluorescence imaging system. According to the
temporal relations between cells and fluorescent embedded into the culture medium, we can
estimate the cell- substratum traction and shear force.
Third: Fluorescent stains for transcription factors of the slug, snail, β-catenin, and p-120
will be done in cells respectively at the ambient pressure and at the pressure of 635 mmHg for
12 hours. Western blots will be performed for those have different presentations. The
wounded cells were incubated at the atmospheric pressure for 12 hours after the knockdown
of slug and snail. We wish to compare the wound-healing process and E-cadherin expressions
in cells after the above treatment with those at the pressure of 635 mmHg.
Project IDs
Project ID:PC10308-0981
External Project ID:MOST103-2314-B182-003
External Project ID:MOST103-2314-B182-003
Status | Finished |
---|---|
Effective start/end date | 01/08/14 → 31/07/15 |
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