Parallel probing of drug uptake of single cancer cells on a microfluidic device

Abstract

Drug resistance is frequently developing during treatment of cancer patients. Intracellular drug uptake is one of the important characteristics to understand mechanism of drug resistance. However, the heterogeneity of cancer cells requires the investigation of drug uptake at the single cell level. Here, we developed a microfluidic device for parallel probing of drug uptake. We combined a v-type valve and peristaltic pumping to select individual cells from a pool of prostate cancer cells (PC3) and place them successively in separate cell chambers in which they were exposed to the drug. Six different concentrations of doxorubicin, a naturally fluorescent anti-cancer drug, were created in loop-shaped reactors and exposed to the cell in closed 2 nL volume chambers. Monitoring every single cell over time in 18 parallel chambers revealed increased intracellular fluorescence intensity according to the dose of doxorubicin, as well as nuclear localization of the fluorescent drug after 2 h of incubation. The herein proposed technology demonstrated a first series of proof of concept experiments and it shows high potential to use for probing drug sensitivity of single cancer cell.

High-Throughput Methods in the Discovery and Study of Biomaterials and Materiobiology

“… 626 The development of fabrication techniques to produce polymeric devices in the late 1990s 627 opened a new paradigm in micrototal analysis systems (μTAS) and lab-on-a-chip (LOC) technology, especially for biological applications including chemotaxis, 628 − 630 cell biophysics, 631 , 632 DNA sequencing, 633 − 635 and cell culture assay. 636 − 638 Besides benefits from a small volume, for example, low sample consumption, fast analysis, and low cost, microfluidics offers possibilities for high-throughput and combinatorial analysis by parallelization, automation, and integration with gradient generators, 639 , 640 , 649 , 650 , 641 − 648 sensors, 651 , 652 and actuators. 597 , 653 , 662 , 663 , 654 − 661 Also, microfluidic devices coupled with various micro/nano-patterning 658 , 661 and surface modification techniques 414 enabled to mimic and study fundamental cellular processes, including cell adhesion, proliferation, and differentiation.…”

“… 705 − 707 Over hundreds of microfluidic single-cell capture units were easily parallelized to increase the throughput capabilities in a reactor with relatively low fabrication cost compared to other techniques, such as a single cell micromanipulator 708 − 710 or optical tweezer. 711 , 712 An integrated channel network allowed further manipulation and analysis on isolated single cells such as cell culture, 713 , 714 drug dose–response, 649 , 715 , 716 and DNA sequencing. 717 − 720 Skelley et al developed a device to trap and pair thousands of cells by adapting microfluidic capture structures ( Figure 30 B(b)) and showed reprogramming in a hybrid between mouse ECSs and mouse embryonic fibroblasts (mEFs) through on-chip electrofusion.…”

“…Microfluidics, a technology that manipulates fluids at small scales (typically from 10 –8 to 10 –18 liters) in narrow (10 –6 to 10 –8 meters) channels, provides engineered and integrated platforms for chemical, biotechnological, and biological analysis applications . The development of fabrication techniques to produce polymeric devices in the late 1990s opened a new paradigm in micrototal analysis systems (μTAS) and lab-on-a-chip (LOC) technology, especially for biological applications including chemotaxis, − cell biophysics, , DNA sequencing, − and cell culture assay. − Besides benefits from a small volume, for example, low sample consumption, fast analysis, and low cost, microfluidics offers possibilities for high-throughput and combinatorial analysis by parallelization, automation, and integration with gradient generators, ,,,,− sensors, , and actuators. ,,,,− Also, microfluidic devices coupled with various micro/nano-patterning , and surface modification techniques enabled to mimic and study fundamental cellular processes, including cell adhesion, proliferation, and differentiation. Recent technical development in multilayer device fabrication to construct 3D models offered new tools that resemble complex microenvironments in nature for studying physiological processes. − In this chapter, section will review microfluidic HTS approaches that have been developed to improve efficiency and accuracy of data collection and analysis, and section will outline microfluidic systems to recreate physicochemical stimuli and physiological/biomimetic microenvironments for screening cell–material interactions.…”

Chips for Biomaterials and Biomaterials for Chips: Recent Advances at the Interface between Microfabrication and Biomaterials Research

“…[10] This allows single cells to be isolated and analyzed, allowing the behavior of heterogeneous populations of cells to be better understood. [11,12] The same microfabrication techniques used to produce chips for mixing, separation and analysis have also been used to produce on-chip systems for cell culture. So-called "organ-on-chip" environments mimic the cell culture conditions in a biological tissue or organ, which allows the reduction or replacement of animal studies.…”

A valve‐based microfluidic device for on‐chip single cell treatments

“…Due to inherent drawbacks of the above methods, droplet microfluidic technique has been developed as a useful approach to conduct single‐cell analysis . Cells are isolated into monodispersed nanoliter droplets whose large surface‐volume ratio significantly increases the efficiency of assays.…”