Morphological Evaluation of Nonlabeled Cells to Detect Stimulation of Nerve Growth Factor Expression by Lyconadin B

Abstract

The success of drug development is greatly influenced by the efficiency of drug screening methods. Recently, phenotypebased screens have raised expectations, based on their proven record of identifying first-in-class drugs at a higher rate. Although fluorescence images are the data most commonly used in phenotype-based cell-based assays, nonstained cellular images have the potential to provide new descriptive information about cellular responses. In this study, we applied morphology-based evaluation of nonlabeled microscopic images to a phenotype-based assay. As a study case, we attempted to increase the efficiency of a cell-based assay for chemical compounds that induce production of nerve growth factor (NGF), using lyconadin B as a model compound. Because the total synthesis of lyconadin B was accomplished very recently, there is no well-established cell-based assay scheme for further drug screening. The conventional cell-based assay for evaluating NGF induction requires two types of cells and a total of 5 days of cell culture. The complexity and length of this assay increase both the risk of screening errors and the cost of screening. Our findings show that analysis of cellular morphology enables evaluation of NGF induction by lyconadin B within only 9 h.

Morphology-Based Analysis of Myoblasts for Prediction of Myotube Formation

“…We have reported the efficient application of this method to the quality assessment of mesenchymal stem cells 6,7 and iPSCs 8 and to the screening of PC12 cells. 9 In this work, we demonstrate that our morphology-based cellular evaluation method can be applied to myotube formation assays for drug screening. As a model case, we have evaluated the cellular responses of C2C12 cells, which form multinuclei cells and acquire morphological features of differentiating muscle cells.…”

“…It is a fact that whether we can robustly apply our methodology to other types of myoblasts is the next important question to address in detail. Nevertheless, our morphological analysis concept based on statistical features of groups of cells has been successfully applied to a variety of cell types (mesenchymal stem cells, 6,7 adrenal gland cells, 9 and other cells [PCT Patent WO2011021391A1]), including different lots. Therefore, we consider the applicability of our methodology to be broad.…”

“…We have reported the efficient application of this method to the quality assessment of mesenchymal stem cells 6,7 and iPSCs 8 and to the screening of PC12 cells. 9…”

Label-free morphological sub-population cytometry for sensitive phenotypic screening of heterogenous neural disease model cells

“…Label-free phenotypic analysis yields more information on live and intact cells than labeled phenotypic analysis [4][5][6][7] . The advantages of label-free image analysis are high-throughput screening, low cost, and non-invasive cell evaluation [7][8][9][10][11][12][13][14][15][16] . Additionally, label-free imaging enables the acquisition of transitional cellular information-during the culture period-instead of only end-point information.…”

“…Conventional label-free image analysis utilizes two types of image data utilization concepts. The first type involves the extraction of multiple features from the cells in the image to describe the morphological features of cells (area, roundness, and peripheral features 7,[10][11][12][14][15][16] ), intensity patterns, and textures 10,11,14,[16][17][18][19] , which reflect the morphological characteristics observed in microscopy. Therefore, analysis results, especially the validity of the trained model, can be biologically interpreted.…”