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Investigating Cancer Microtissue Metabolism by Measuring Glucose and Lactate Levels in a Hanging Drop Network
Fabricating hanging drop biosensors, forming 3D cancer microtissues of various sizes, and measuring their glucose and lactate metabolism lets us study diffusion and uptake kinetics in 3D microtissues.
Keywords: 3D microtissues
Cancer
Metabolism
Warburg effect
Microfluidics
Hanging-drop network
Biosensors
Diffusion-reaction modelling
Microtissues (MTs) are three-dimensional (3D) cell aggregates that feature more organ-like behavior than traditional two-dimensional cell cultures. With the appropriate cell type and source, these 3D MTs can model any healthy (e.g. liver, pancreas, heart, etc.) or diseased (e.g. fatty liver, tumors, etc.) organ. An important feature of 3D MTs is their ease of transfer into so-called microphysiological systems that interconnect different organ models. However, to justify this increase in system complexity, it is necessary to quantify how 3D MTs offer an advantage over conventional models. We do this by elucidating the heterogeneous metabolism caused by diffusion and consumption processes within 3D MTs.
In this project, we use a simple 3D MT cancer model using a human colon cancer cell line. Cells are aggregated in a sphere and can be used to investigate cancer metabolism. Glucose metabolism within cancer cells features a high level of aerobic glycolysis. This process, known as the Warburg effect, causes a build-up of lactate in the cancer microenvironment. When glucose is depleted, cancer cells consume the lactate as an alternate carbon source. Since this process is concentration-dependent, it is the perfect candidate to elucidate the heterogeneous metabolism in 3D MTs.
These glucose and lactate concentrations can be measured for single cancer microtissues with the cutting-edge integrated biosensors developed at the Bio Engineering Laboratory. These biosensors feature an enzyme-based hydrogel that is sensitive to very low concentrations of nutrients. Ultimately, treating cancer MTs with compounds will allow to investigate their metabolism, starvation dynamics, diffusion/consumption processes, and the Warburg effect.
Microtissues (MTs) are three-dimensional (3D) cell aggregates that feature more organ-like behavior than traditional two-dimensional cell cultures. With the appropriate cell type and source, these 3D MTs can model any healthy (e.g. liver, pancreas, heart, etc.) or diseased (e.g. fatty liver, tumors, etc.) organ. An important feature of 3D MTs is their ease of transfer into so-called microphysiological systems that interconnect different organ models. However, to justify this increase in system complexity, it is necessary to quantify how 3D MTs offer an advantage over conventional models. We do this by elucidating the heterogeneous metabolism caused by diffusion and consumption processes within 3D MTs.
In this project, we use a simple 3D MT cancer model using a human colon cancer cell line. Cells are aggregated in a sphere and can be used to investigate cancer metabolism. Glucose metabolism within cancer cells features a high level of aerobic glycolysis. This process, known as the Warburg effect, causes a build-up of lactate in the cancer microenvironment. When glucose is depleted, cancer cells consume the lactate as an alternate carbon source. Since this process is concentration-dependent, it is the perfect candidate to elucidate the heterogeneous metabolism in 3D MTs.
These glucose and lactate concentrations can be measured for single cancer microtissues with the cutting-edge integrated biosensors developed at the Bio Engineering Laboratory. These biosensors feature an enzyme-based hydrogel that is sensitive to very low concentrations of nutrients. Ultimately, treating cancer MTs with compounds will allow to investigate their metabolism, starvation dynamics, diffusion/consumption processes, and the Warburg effect.
During this master thesis project, the student will:
1. assist in the fabrication of the cutting-edge integrated biosensors with a microfluidic hanging-drop network;
2. form cancer MTs of varying sizes to serve as the glucose-consuming organ;
3. use the hanging-drop biosensor in order to monitor the metabolism of cancer MTs;
4. implement a data analysis pipeline to extract diffusion/consumption parameters from the biosensor;
5. further disturb glucose metabolism by including glucose-transporter-inhibitors.
Completing these tasks will help in establishing advanced cancer microtissue metabolism models.
The student will learn:
- Principles of cleanroom microfabrication:
o Microelectrode fabrication
o SU8 processes
- Soft lithography / PDMS fabrication techniques
- Electrode functionalization
- Advanced open space microfluidics design and operation
- Microscopy
- Mammalian 3D cell cultures – 3D microtissue formation and handling
- Metabolism parameter extraction
During this master thesis project, the student will:
1. assist in the fabrication of the cutting-edge integrated biosensors with a microfluidic hanging-drop network;
2. form cancer MTs of varying sizes to serve as the glucose-consuming organ;
3. use the hanging-drop biosensor in order to monitor the metabolism of cancer MTs;
4. implement a data analysis pipeline to extract diffusion/consumption parameters from the biosensor;
5. further disturb glucose metabolism by including glucose-transporter-inhibitors.
Completing these tasks will help in establishing advanced cancer microtissue metabolism models.
The student will learn:
- Principles of cleanroom microfabrication:
o Microelectrode fabrication
o SU8 processes
- Soft lithography / PDMS fabrication techniques
- Electrode functionalization
- Advanced open space microfluidics design and operation
- Microscopy
- Mammalian 3D cell cultures – 3D microtissue formation and handling