An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening

Abstract

Background Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer treatment. Elucidating the cytotoxicity of anticancer drugs at single-cell level in a high-throughput way is thus of great value for developing precision therapy. However, current techniques suffer from limitations in dynamically characterizing the responses of thousands of single cells or cell clones presented to multiple drug conditions. Methods We developed a new microfluidics-based “SMART” platform that is Simple to operate, able to generate a Massive single-cell array and Multiplex drug concentrations, capable of keeping cells Alive, Retainable and Trackable in the microchambers. These features are achieved by integrating a Microfluidic chamber Array (4320 units) and a six-Concentration gradient generator (MAC), which enables highly efficient analysis of leukemia drug effects on single cells and cell clones in a high-throughput way. Results A simple procedure produces 6 on-chip drug gradients to treat more than 3000 single cells or single-cell derived clones and thus allows an efficient and precise analysis of cell heterogeneity. The statistic results reveal that Imatinib (Ima) and Resveratrol (Res) combination treatment on single cells or clones is much more efficient than Ima or Res single drug treatment, indicated by the markedly reduced half maximal inhibitory concentration (IC50). Additionally, single-cell derived clones demonstrate a higher IC50 in each drug treatment compared to single cells. Moreover, primary cells isolated from two leukemia patients are also found with apparent heterogeneity upon drug treatment on MAC. Conclusion This microfluidics-based “SMART” platform allows high-throughput single-cell capture and culture, dynamic drug-gradient treatment and cell response monitoring, which represents a new approach to efficiently investigate anticancer drug effects and should benefit drug discovery for leukemia and other cancers.

Optical Detection of Cancer Cells Using Lab-on-a-Chip

“…Taking advantage of microfluidics, it was possible to separate single cells and promote the formation of single-cell-derived colonies for drug testing. An array with more than 4000 microchambers coupled to a concentration gradient generator allowed researchers to distinguish cell heterogeneity of both a leukemia cell line and primary tumor cells [ 30 ]. The device enables researchers to monitor cell viability over several time points by registering the fluorescence of live and dead cells.…”

Advances in organ‐on‐a‐chip for the treatment of cardiovascular diseases

“…168 Remarkably, Ooc technology could simulate the concentration gradient distribution of drugs in the body through a drug gradient concentration control system to assess the toxicity and effects of drugs at different concentrations. 169,170 For example, Grant and colleagues developed a PDMS organ chip containing two parallel channels separated by a porous membrane arranged by living cells and described a combined experimental and computational strategy to estimate changes in drug concentration over time without prior knowledge of the partition or diffusion coefficients. The application of this strategy not only increases the usefulness of organoids in clinical research projects, but also allows their application to other forms of drug testing.…”

“…On the one hand, some cellular biological responses are dependent on specific biomolecular concentrations and thus the control of drug concentration gradients is of great significance in drug toxicity screening 168 . Remarkably, Ooc technology could simulate the concentration gradient distribution of drugs in the body through a drug gradient concentration control system to assess the toxicity and effects of drugs at different concentrations 169,170 . For example, Grant and colleagues developed a PDMS organ chip containing two parallel channels separated by a porous membrane arranged by living cells and described a combined experimental and computational strategy to estimate changes in drug concentration over time without prior knowledge of the partition or diffusion coefficients.…”

Mapping the Potential of Microfluidics in Early Diagnosis and Personalized Treatment of Head and Neck Cancers

“…Overcoming the challenge of increasing the sensitivity and specificity of microfluidic technology with regard to the screening and detection of key biomarkers of HNC requires a large testing sample population to reliably translate these techniques into a clinical setting [141]. High-throughput screening (HTS) is a technique that works by testing and analyzing large numbers of chemical or biological compounds against specific targets in a very short time, with the goal to develop either drug or diagnostic tools [142,143]. Researchers have observed that many limitations of existing HTP screening systems can be resolved by using microfluidic technologies since they can be produced at a much lower cost and work on liquid volumes at the level of nano-to microliters or smaller [143].…”