Researchers from the University of Twente have developed a new analysis method to rapidly study millions of individual cells and the proteins they secrete to form tissues. The researchers termed their method Extracellular Protein Identification Cytometry (EPIC). "This changes the way we can study living matter and has many potential uses such as driving fabrication of replacement organs, and accelerating development and testing of medicines”, says researcher Marieke Meteling. Her work has recently been published in the prestigious scientific journal Advanced Materials.
The tissues that make up your organs consist of cells within an extracellular matrix that is produced by the cells. This matrix is not only important for how the cells function but is also an important biomarker for various diseases. Although several techniques exist to analyse this matrix, these approaches are either slow or offer poor resolution, and many require tissue destruction. This has limited the clinical and societal impact of these applications.
UT researchers from the lab of Prof. Leijten combined advanced microfluidic technology and biomaterial science with an existing measurement technique named flow cytometry to allow for the world-first high-throughput, high-resolution screening platform for the quantification of extracellular matrix.
Applications
For the successful engineering of living tissues, creation of disease models, realistic testing of drugs, and development of regenerative medicine, it is essential that cells produce the intended type of extracellular matrix. EPIC can reveal how individual cells create and modify their surroundings and do this for a large number of cells.
“Researchers can now study important questions such as what percentage of cells is affected by a drug, and why certain cells are sensitive or not to a specific treatment”, explains Meteling. EPIC can provide new insights into the tissue formation process, or how tissue destruction by specific cells occurs or is altered by treatments. These insights could advance treatments for diseases like osteoarthritis, fibrosis, and cancer.
Engineered 3D microenvironments
EPIC uniquely combines microfluidic technology, biomaterial science, immunolabelling, and flow cytometry. Microfluidic devices produce millions of tiny droplets that each contain a single cell. These microscopic droplets are then solidified into miniature hydrogel spheres in which each cell can be cultured for several weeks to deposit and/or deconstruct the extracellular matrix.
Fluorescence antibodies that bind specifically to one type of matrix proteins are then used to show how each cell behaves. These markers can then be measured in high-throughput using flow cytometry. Flow cytometry is a powerful tool for any field that makes use of living cells. Until now, flow cytometry couldn’t be used to measure matrix proteins because extracting the cells required the use of enzymes that severely damaged or removed the matrix.
The UT researchers solved this problem by enabling cells to build or remodel matrix directly inside miniature 3D microgels. These microgels can go straight into a flow cytometer without damaging the matrix. This method uniquely maintains all matrix and cellular information for measurement. The non-destructive nature of the approach makes it possible to measure and isolate (individual) cells of interest, which can then be used for other high-content analyses such as confocal imaging, genomics, spatial proteomics, and mass spectroscopy.
More information
The research was conducted by Marieke Meteling under the supervision of Prof Dr Jeroen Leijten, both members of the Leijten lab, in the Developmental BioEngineering group (Faculty of S&T) of the BioEngineering Technologies department at the University of Twente. The paper “High-Throughput Single-Cell Analysis of Local Nascent Protein Deposition in 3D Microenvironments via Extracellular Protein Identification Cytometry (EPIC)” by Marieke Meteling, Castro Johnbosco, Alexis Wolfel, Francisco Conceição, Kannan Govindaraj, Liliana Moreira Teixeira, and Jeroen Leijten, was recently published in Advanced Materials.
