Develop automated, high-throughput organ-on-a-chip cell models using high-content imaging and 3D analysis
What is organ-on-a-chip?
Organ-on-a-chip (OoC) is a technology that uses microfabrication techniques to create miniature models of biological organs, such as the lung, heart, or gut, on a chip-sized device. These microfabricated devices are made up of living cells that are grown on a microscale platform and mimic the structure and function of the organ they represent. The cells are typically arranged in a way that mimics the native three-dimensional structure of the organ and is perfused with fluids, such as blood or air, to represent the physiological environment of the organ.
OoC technology is used to create more accurate and reliable models of organs and tissues that can better replicate the complex microenvironment and interactions of cells within an organ. 3D cell models can be used to study disease, drug development, and toxicology in a more accurate and realistic way than traditional 2D cell cultures.
The OrganoPlate® 3-lane 64 culture chip and schematic representation along with the an Illustration of a tubule of cells grown against an ECM gel.
How does organ-on-a-chip technology work
Organ-on-a-chip technology typically consists of a polymer material that is molded into the shape that mimics some aspects of morphology of the organ of interest. Cells are then seeded onto the chip and allowed to grow and form functional 3D structures that would resemble cell composition and structure of tissues. In some cases, the chip may be designed to include microfluidic channels mimicking the microvasculature of an organ to provide blood flow or other physiological conditions including nutrients and oxygen to the cells.
In order to have a more realistic representation of the organ of interest, various cell types can be combined to form a 3D structure, this can be done by using different layer of cells, or by using hydrogel-based matrices to mimic the extracellular matrix of the organ. Various techniques can be applied to mimic the mechanical, electrical, and chemical microenvironment of the organ. For instance, the chip can be perfused with fluids to provide blood flow, or it can be mechanically stimulated to mimic heart contractions. Additionally, sensors can be integrated into the chip to measure things like oxygen, pH, and temperature, in order to monitor the health and function of the cells.
The chip is placed in an incubator where cell growth can be monitored using various techniques such as microscopy, imaging, or biochemical assays. Once the chip is fully functional, scientists can use it to study disease, drug development, and toxicology in a controlled and highly reproducible way. This is because the chip can be used to mimic the same conditions, in the same way, every time an experiment is run, allowing scientists to compare data consistently across different experiments and treatments.
Automation of the organ-on-a-chip assay for high-throughput screening
Here we describe a workflow for automation of OoC culture, as well as monitoring, and automated cell analysis. The automated method utilizes an integrated work-cell comprising several instruments that allow the automation and monitoring of cell culture. The high-content imaging system enables the characterization of 3D cell model development, as well as testing the effects of compounds. The integrated system includes the ImageXpress® Micro Confocal High-Content Imaging System, an automated CO2 incubator, a liquid handler (Biomek i7), and collaborative robot. We developed methods for automation of cell seeding, media exchange, and for monitoring the development and growth of 3D vasculature. In addition, the method facilitates automated compound testing and evaluation of toxicity effects.
Watch OoC poster presentation with Oksana Sirenko, Sr. Application Scientist on how our high-content imaging solutions can scale up and automate 3D imaging of organ-on-a-chip systems.
Figure 1. Layout of the individual instruments in the workcell is illustrated in (A). The instruments are controlled by an integrated software (Green Button Go) that allows for set up of processes. An example of the process to monitor cells in culture is shown in (B). Here, the plates are moved from the incubator to the ImageXpress Confocal HT.ai for imaging in brightfield and then back to the incubator. The process can also be scheduled, and plates that need to be imaged can be entered as a list to enable easier batch processing. More complex routines that include the liquid handler for media exchanges (feeding) can also be implemented.
Organ-on-a-Chip applications and assays
Combining this complex biology with advanced high-content imaging techniques and AI/machine learning 3D analysis capabilities opens up a whole new level of assays. Here we share our methods for automation of the cell culture, assays, and analysis can provide the tools necessary to facilitate and scale up the use of organ-on-a-chip systems.