Differentiating a stem cell into a neuron or glial cell is typically a time-consuming bottleneck in human neurological dysfunction research. In addition, precisely controlling the purity of subtypes in a mixed neuron population can be a complex task. Purity is critical to improve assay reproducibility and eliminate unwanted cell types that may negatively impact results.
At the University of Minnesota’s Stem Cell Business Incubator in Minneapolis, Anatomi Corporation has commercialized a technology, known as the Chrono™ platform. The technology eliminates these bottlenecks so that researchers can focus on their research goals.
Using the Chrono™ platform, culture media formulations containing small molecules, growth factors, and morphogens rapidly direct differentiation of human induced pluripotent stem cells (hiPSCs) into specific neuronal subpopulations. This process is completed in about seven days, which is significantly shorter compared to traditional methods that take 30 days or longer to complete.
Time is of the essence in neurobiology research and is also the driving principle of the Chrono platform. As the Anatomi CEO, Patrick Walsh, points out, “An accelerated pace of manufacturing means that experiments are completed faster.” He continues, “On top of that, simplified protocol development and a workflow that is easily managed by users of any skill level enable researchers to spend more time interpreting data rather than preparing experiments.”
In addition to considerable time efficiencies, Walsh notes that the Chrono platform also enhances process reliability and reproducibility.
“Our first product is a sensory neuron, and we’re rapidly expanding our portfolio to include cortical glutamatergic neurons, striatal GABAergic neurons, Schwann cells, and more,” says Vincent Truong, Chief Operating Officer at Anatomi. “Using the Chrono platform, we can develop thousands of the specific neuronal subtypes found in the human body.”
Undifferentiated cells go through various stages of development; each offering landmark gene expression. Therefore, Anatomi relies heavily on immunocytochemistry assays to quantify populations and determine the purity of a final product during product and application development. This is where two Cytation 1 cell imaging multimode readers, housed at the University’s Laboratory for Stem Cell Automation, are essential to the process.
Cytation 1 increases throughput when screening factors that show differentiation propensity. The Gen5 software facilitates rapid data analysis. Cytation 1 is equipped with Agilent BioTek CO2 Gas Controller to regulate the environment throughout the experimental timeframe. In use, Cytation 1 acquires a four-channel image of cells stained with three different markers. The DAPI channel provides a whole cell number. The antigen of choice is selected using the other imaging channels and expressed as a percentage of that whole cell number. This process helps to guide downstream iterations and experiments.
The Anatomi team is taking full advantage of Cytation 1 and Gen5, including the ability to show differentiation processes in accelerated time over the entire experimental timeframe. Truong explains, “It’s impractical to gather time-lapse data in traditional bulky workflows, but with our short protocol and Cytation 1, we can capture really interesting and informative time-lapse data.”
The robust Cytation 1 is a welcome part of Anatomi’s process. As Walsh surmises, “It’s not just efficient, it’s liberating. We can set Cytation 1 up and attend to other tasks while it runs, and we’re very confident in the data that it generates.”