Rapid Detection of Direct Compound Toxicity and Trailing Detection of Indirect Cell Metabolite Toxicity in a 96-Well Fluidic Culture Device for Cell-Based Screening Environments: Tactics in Six Sigma Quality Control Charts.

Bob Lubamba,Randall Mcclelland,Timothy Jensen

doi:10.3390/app12062786

Abstract

Microfluidic screening tools, in vitro, evolve amid varied scientific disciplines. One emergent technique, simultaneously assessing cell toxicity from a primary compound and ensuing cell-generated metabolites (dual-toxicity screening), entails in-line systems having sequentially aligned culture chambers. To explore dual-tox screens, we probe the dissemination of nutrients involving 1-way transport with upstream compound dosing, midstream cascading flows, and downstream cessation. Distribution of flow gives rise to broad concentration ranges of dosing compound (0→ICcompound100) and wide-ranging concentration ranges of generated cell metabolites (0→ICmetabolites100). Innately, single-pass unidirectional flow retains 1st pass informative traits across the network, composed of nine interconnected culture wells, preserving both compound and cell-secreted byproducts as data indicators in each adjacent culture chamber. Thereafter, to assess effective compound hepatotoxicity (0→ECcompound100) and simultaneously classify for cell-metabolite toxicity (0→ECmetabolite100), we reveal utility by analyzing culture viability against ramping exposures of acetaminophen (APAP) and nefazodone (NEF), compounds of hepatic significance. We then discern metabolite generation with an emphasis on amplification across μchannel multiwell sites. Lastly, using conventional cell functions as indicator tools to assess dual toxicity, we investigate a non-drug induced liver injury (non-DILI) compound and DILI compound. The technology is for predictive evaluations of new compound formulations, new chemical entities (NCE), or drugs that have previously failed testing for unresolved reasons.

Highlights

New cell culture devices are populating life science and pharmaceutical R&D benches at escalating rates, each with unique features and various functions [1–3]
The multiwell device, unidirectional flow, and six-sigma analysis presented here demonstrate the rapid detection of direct compound toxicity and trailing detection of cellbyproduct influencers across a cell culture platform
The unmasking of cell-byproduct mechanisms allow for evaluations of metabolite-mediated toxicity through slow, medium, and fast-acting toxicity profiles, formerly concealed

Summary

Introduction

New cell culture devices are populating life science and pharmaceutical R&D benches at escalating rates, each with unique features and various functions [1–3]. Efforts in development are driven by the need to improve human-relevant experimental systems earlier in the drug evaluation process. In regard to platform physical parameters, μfluidic configurations commonly include a culture chamber, or multiple linked chambers, nutrient flow, and controllers such as pumps, valves, and fluid transport tubes [10]. While such devices can be skillfully prototyped, the refinement into mass manufacturing of product has tangible engineering constraints. End-user objectives align with targeted use-model strategies, needs for lab compatible accessories, and design conditions that prompt development constraints

Methods

Discussion

Conclusion