Optimization of Live Cell Confocal Imaging Bioreactor for Mechanical Strain Application

Abstract

The human body is put under physical strain on a daily basis, especially in bones. Therefore, learning how mechanical strain affects bone cells, and more importantly, the mesenchymal stem cells (MSCs) that make them, is crucial to understanding the changes in structure and function that can occur.

To simulate mechanical stress, a “strain bioreactor” is used to create a vacuum on a membrane which stretches it. The vacuum can apply various levels of strain to the membrane. This needs to be calibrated to ensure the appropriate magnitude of strain is evenly distributed across the areas of interest on the membrane. Using fluorescent beads, I tested if repeated use of one membrane affected the quality of data collected over time. Next, I used new membranes to test whether or not 2, 4, or 6% strain was accurately being applied to various locations along the center of the membrane.

Confirmation that the correct level of strain is applied to membranes is vital for future live cell experiments. Without understanding how strain is being applied, future research could improperly collect data and find inaccurate results. Instead, assessments on strain application ensure this does not occur. This allows for important research to be conducted on how mechanical strain on MSCs relate to associated diseases and how to improve their treatment.

This document is currently not available here.

Share

COinS
 

Optimization of Live Cell Confocal Imaging Bioreactor for Mechanical Strain Application

The human body is put under physical strain on a daily basis, especially in bones. Therefore, learning how mechanical strain affects bone cells, and more importantly, the mesenchymal stem cells (MSCs) that make them, is crucial to understanding the changes in structure and function that can occur.

To simulate mechanical stress, a “strain bioreactor” is used to create a vacuum on a membrane which stretches it. The vacuum can apply various levels of strain to the membrane. This needs to be calibrated to ensure the appropriate magnitude of strain is evenly distributed across the areas of interest on the membrane. Using fluorescent beads, I tested if repeated use of one membrane affected the quality of data collected over time. Next, I used new membranes to test whether or not 2, 4, or 6% strain was accurately being applied to various locations along the center of the membrane.

Confirmation that the correct level of strain is applied to membranes is vital for future live cell experiments. Without understanding how strain is being applied, future research could improperly collect data and find inaccurate results. Instead, assessments on strain application ensure this does not occur. This allows for important research to be conducted on how mechanical strain on MSCs relate to associated diseases and how to improve their treatment.