Time to Accelerate the Pace of Neurobiology Research
University of Minnesota Stem Cell Incubator, Anatomi Corporation
Differentiating a stem cell into a neuron or glial cell is typically a time-consuming bottleneck in human neurological dysfunction research. On top of this, precisely controlling the purity of sub-types in a mixed neuron population can be a complex task, yet 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, to eliminate this bottleneck so that researchers can focus on their research goals.
Using the patented 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 when it comes to neurobiology research and is also the driving principle of the Chrono Platform. As Anatomi’s 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 Multi-Mode 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; its Gen5™ Software facilitates rapid data analysis. One Cytation is equipped with BioTek’s 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, then the antigen of choice is selected using the other imaging channels and expressed as a percentage of that whole. This 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.”
Live time course analysis of passage-free differentiation of hiPSCs into spinal neurons demonstrating spontaneous action potentials by day ten based on calcium imaging. Day timer shown at bottom right of video. Final video frames indicate the field of view fixed and stained for beta-III tubulin and DAPI to indicate commitment to spinal neurons. Performed in collaboration with Dr. James Dutton at the Stem Cell Institute, University of Minnesota.
Anatomi Chrono™ sensory neurons exhibit extensive axon outgrowth and cluster in ganglion-like structures. Sensory neurons are extensively characterized by immunofluorescence staining and show high expression for peripheral neuron markers (Peripherin, BRN3A, ISL1-green), and general neuronal marker (TUJ1-red).
To learn more about ANATOMI, visit their web site.
Thanks to Anatomi’s CEO, Patrick Walsh for sharing his BioTek experience.