One Hour In Vivo-like Phenotypic Screening System for Anti-cancer Drugs Using a High Precision Surface Plasmon Resonance Device

Abstract

In anti-cancer drug (candidate) screening, there is the need for evaluation at physiological concentrations similar to in vivo. This is often performed by three-dimensionally (3D) cultured cells; however, it requires a long culture period of 2-4 weeks with tedious experimental procedures. Here, we report on a high precision surface plasmon resonance (HP-SPR)-3D system. We developed the system with average fluctuation of 50 ndeg s-1 using two-dimensionally cultured cells attached onto a sensor chip by applying collagen on the top to change their activity into in vivo-like conditions without cell division. It allowed in vivo-like phenotypic screening for anti-cancer drugs within 1 h of drug addition. The data were collected as the stable linear signal change parts for at least 5 min after 25 min following drug addition. The results provided compatibility to clinically related chemosensitivity test for anti-cancer (P <0.001) using two cell lines of pancreatic cancer and three anti-cancer drugs to represent differences in individual gene expression and drug mode of action.

Rapid and synchronized dormancy-broken Kyoho grape seed endosperm without extraction or concentration shows significant effect on both anti-cancer and pro-immunity

“…Cell based assay was performed by using our proprietary High Precision-Surface Plasmon Resonance (HP-SPR)-3D system, non-label and -invasive 1h phenotypic screening by mitochondrial membrane potential, as follows [13]. Twodimensionally cultured viable cells were self-adhered onto an HP-SPR sensor chip, and then collagen was overlaid to obtain in vivo-like cell status.…”

“…The cell response change was measured for about 1h after the sample addition. The Collagen gel droplet embedded culture drug sensitivity test (CD-DST) was also used as a 3D cell culture method [13,14]. Cancer cell lines for human were pancreas MIA-PaCa2, breast MCF-7 and liver Hep G2, and that for canine breast was SNP.…”

Liver mitochondrial activation and fat-lowering effects of rapid and synchronized dormancy-breaking ginger rhizome assumed to be administered orally

“…Cell based assay was performed by using our proprietary High Precision-Surface Plasmon Resonance-Three dimension (HP-SPR-3D) system, non-label andinvasive 1h phenotypic screening by mitochondrial membrane potential, as follows [13]. Two-dimensionally cultured viable cells were self-adhered onto an HP-SPR sensor chip, and then collagen was overlaid to obtain in vivo-like cell status.…”

A Facile Assay of Epithelial-mesenchymal Transition Based on Cooperativity Quantification of Cellular Autonomous Motions

“…1 Especially, the loss of cohesiveness to surrounding cells and increased motility of epithelial cancer cells are critical steps for invasive cancer and metastasis, therefore, EMT is becoming a primary target of anticancer drugs. 2,3 Either for the rapid screening of lead compounds or for the precise evaluation of the potency and efficacy of their structural derivatives, the methods to quantify the EMT progression are of essential importance. However, complex molecular changes during EMT 4 make it difficult to evaluate EMT by isolated molecular targets, like receptors and enzymes, as commonly used for other types of drug screening.…”