A Microfluidic Device for Long‐Term Maintenance of Organotypic Liver Cultures

Abstract

Liver cultures may be used for disease modeling, testing therapies, and predicting drug‐induced injury. The complexity of the liver cultures has evolved from hepatocyte monocultures to co‐cultures with non‐parenchymal cells and finally to precision‐cut liver slices. The latter culture format retains liver's native biomolecular and cellular complexity and therefore holds considerable promise for in vitro testing. However, liver slices remain functional for ≈72 h in vitro and display limited utility for some disease modeling and therapy testing applications that require longer culture times. This paper describes a microfluidic device for longer‐term maintenance of functional organotypic liver cultures. This microfluidic culture system is designed to enable direct injection of liver tissue into a culture chamber through a valve‐enabled side port. Liver tissue is embedded in collagen and remained functional for up to 31 days, highlighted by continued production of albumin and urea. These organotypic cultures also express several enzymes involved in xenobiotic metabolism. Conversely, matched liver tissue embedded in collagen in a 96‐well plate loses its phenotype and function within 3–5 days. The microfluidic organotypic liver cultures described here represent a significant advance in liver cultivation and may be used for future modeling of liver diseases or for individualized liver‐directed therapies.

Microfluidic Organoid Cultures Derived from Pancreatic Cancer Biopsies for Personalized Testing of Chemotherapy and Immunotherapy

“…Fabrication of Microfluidic Devices: Two types of microfluidic devices (see Figures 2A and 4A) were fabricated using standard soft lithography techniques. [17,46] Device 1 was designed to organize single cells into organoids whereas device 2 was used to seed intact organoids. The detailed protocol for fabricating molds and PDMS devices is described in the Supplementary Information.…”

“…Operation of a device to open the valve was described recently. [ 46 ] Briefly, a device was connected to a vacuum line and placed on a microscope in a biosafety hood. The valve was opened by negative pressure to allow insertion of 27‐gauge needle and transfer of organoids into the culture chamber containing in microwells.…”

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“…[ 91–96 ] Microfluidic systems provide a special culture environment that will influence cells and tissue growth states by confining relatively large numbers of cells to a small area. [ 97 ]…”

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