Abstract
The xCELLigence real-time cell analysis (RTCA) system has the potential to detect cellular proliferation, migration, cytotoxicity, adherence, and remodeling. Although the RTCA system is widely recognized as a noninvasive and efficient tool for real-time monitoring of cellular fate, it cannot describe detailed cell morphological parameters, such as length and intensity. Transforming growth factor beta(TGF-β) induced the epithelial–mesenchymal transition (EMT), which produces significant changes in cellular morphology, so we used TGF-β to treat A549 epithelial cells in this study. We compared it with lipopolysaccharide (LPS) and cigarette smoke extract (CSE) as stimulators. We developed an efficient algorithm to quantify the morphological cell changes. This algorithm is comprised of three major parts: image preprocessing, Hough transform (HT), and post-processing. We used the RTCA system to record the A549 cell index. Western blot was used to confirm the EMT. The RTCA system showed that different stimulators produce different cell index curves. The algorithm determined the lengths of the detected lines of cells, and the results were similar to the RTCA system in the TGF-β group. The Western blot results show that TGF-β changed the EMT markers, but the other stimulator remained unchanged. Optics-based computer vision techniques can supply the requisite information for the RTCA system based on good correspondence between the results.
Highlights
Morphological properties are used to distinguish different kinds of cells or the same cell in different stages to help with understanding molecular cell fate [1]
Morphological variation is of the vital parameters that contributes to the change in the cell index and provides a rational approach one of the vital parameters that contributes to the change in the cell index and provides a rational to evaluate the epithelial–mesenchymal transition (EMT) process using the change in the cell index
We found that a high slope, or the change to an arch structure after cellular fate induced by TGF-β [10]
Summary
Morphological properties are used to distinguish different kinds of cells or the same cell in different stages to help with understanding molecular cell fate [1]. Many studies have demonstrated that treatment with transforming growth factor beta (TGF-β) results in a significant change in cellular morphology; epithelial cells are changed from a typical cobblestone type to slender fibroblast-like mesenchymal cells [2,3]. This process is called epithelial–mesenchymal transition (EMT). Morphological changes are an imperative optical indicator to illustrate the effect of TGF-β in the EMT process. Numerous studies have described changes in cell shape by direct microscopic observations [7,8]
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