Dynamic Culture and Selective Extraction of Target Microbial Cells in Self-Assembled Particle Membrane-Integrated Microfluidic Bioreactor Array

Abstract

Various microfluidic devices have overcome many disadvantages common to conventional bioreactor systems by enabling active manipulation of cell-culture conditions, monitoring of cellular responses in high-throughput mode, and extraction of target cells in a relatively rapid and low-cost manner. However, existing microfluidic devices still have limitations, including the complexity of their operation and a lack of availability of dynamic control of the chemical environment. Here, we present a novel microfluidic bioreactor array device capable of not only the stable and dynamic programing of cell-culture environments but also the selective extraction of target cells. This device comprises 64 microchambers in a 16 × 4 array format, and each microchamber is integrated with a robust and nanoporous membrane on one side and an H-shaped entrance on the other. The membrane made of self-assembled particles allowed continuous and sequential delivery of various nutrients containing gene inducers to compartmentalized microbial cells, thereby enabling dynamic cell culturing. Additionally, the H-shaped entrance was used for local and selective blocking of the microchamber by employing UV-curable material, thereby enabling the retrieval of target cells from the device while sequestering nontarget cells in the microchambers. Our results demonstrated that the targeted rare cells could be isolated and separated from a mixture of cells by repeating the extraction procedure. Therefore, we anticipate that this microfluidic bioreactor array device will be widely used for not only screening/extraction but also off-chip postanalyses of various microorganisms.