Investigating Effects of Mechanical Cues on Macrophage Polarization

The skin as an organ is exposed to multiple mechanical stresses (strain, compression, hydrostatic pressure). These stresses are highly relevant both to the development of skin diseases and to finding potential novel therapies. To investigate the role of these different mechanical stresses in skin biology, we have been developing physiologically and mechanically relevant 3D hydrogel models, using cells isolated from patient skin samples thus far focusing on isolated fibroblasts and keratinocytes. While these cell types play important roles in the maintaining homeostasis, following an injury to the skin, a highly orchestrated immune response occurs, involving infiltration of immune cells into damaged tissue from circulation. To further investigate the role of mechanical cues on cells in the dermis during disease and injury, we aim to incorporate immune cells into our 3D hydrogel models to study both the effects of mechanical cues on immune cell phenotype, as well as cross talk with other tissue resident cells in a co-culture model. One of the first immune cell types to infiltrate the damaged dermis are monocytes, which become a source of macrophages. Macrophages play an important role in supporting normal wound closure by clearing debris and coordinating with other resident cell types, such as fibroblasts. If this process is dysregulated, wound closure can be inhibited, as in chronic wounds, or aggravated as in fibrosis. To incorporate macrophages into our hydrogel skin models, we require using unbiased/unpolarized macrophages which and can differentiated from circulatory monocytes using differentiation protocols. These circulatory monocytes can be isolated from patient whole blood using established protocols. The focus of this Masters thesis will be to help establish and validate in house these protocols for i) monocyte isolation ii) macrophage differentiation and iii) macrophage incorporation into our 3D hydrogel skin models.

The skin as an organ is exposed to multiple mechanical stresses (strain, compression, hydrostatic pressure). These stresses are highly relevant both to the development of skin diseases and to finding potential novel therapies. To investigate the role of these different mechanical stresses in skin biology, we have been developing physiologically and mechanically relevant 3D hydrogel models, using cells isolated from patient skin samples thus far focusing on isolated fibroblasts and keratinocytes. While these cell types play important roles in the maintaining homeostasis, following an injury to the skin, a highly orchestrated immune response occurs, involving infiltration of immune cells into damaged tissue from circulation.
To further investigate the role of mechanical cues on cells in the dermis during disease and injury, we aim to incorporate immune cells into our 3D hydrogel models to study both the effects of mechanical cues on immune cell phenotype, as well as cross talk with other tissue resident cells in a co-culture model.

One of the first immune cell types to infiltrate the damaged dermis are monocytes, which become a source of macrophages. Macrophages play an important role in supporting normal wound closure by clearing debris and coordinating with other resident cell types, such as fibroblasts. If this process is dysregulated, wound closure can be inhibited, as in chronic wounds, or aggravated as in fibrosis. To incorporate macrophages into our hydrogel skin models, we require using unbiased/unpolarized macrophages which and can differentiated from circulatory monocytes using differentiation protocols. These circulatory monocytes can be isolated from patient whole blood using established protocols. The focus of this Masters thesis will be to help establish and validate in house these protocols for i) monocyte isolation ii) macrophage differentiation and iii) macrophage incorporation into our 3D hydrogel skin models.

Masters Thesis focused helping establish and validating in house protocols for i) monocyte isolation ii) macrophage differentiation and iii) macrophage incorporation into our 3D hydrogel skin models. Skills required from the student side (all ideal but not necessary): -Cell culture experience -Immunofluorescent Staining and/or Microscopy Skills to be learnt during the project: -Monocyte isolation from human whole blood -Flow cytometry -Macrophage differentiation -3D cell culture -Confocal Imaging -Hydrogel synthesis (if interested) -Western blot

Masters Thesis focused helping establish and validating in house protocols for i) monocyte isolation ii) macrophage differentiation and iii) macrophage incorporation into our 3D hydrogel skin models.

Skills required from the student side (all ideal but not necessary):
-Cell culture experience
-Immunofluorescent Staining and/or Microscopy

Skills to be learnt during the project:
-Monocyte isolation from human whole blood
-Flow cytometry
-Macrophage differentiation
-3D cell culture
-Confocal Imaging
-Hydrogel synthesis (if interested)
-Western blot

Email: jmayner@ethz.ch Please email me, sharing your CV and transcript, if you are interested in the position so we can schedule a meeting. Don't hesitate to reach out if you have any questions!

Email: jmayner@ethz.ch
Please email me, sharing your CV and transcript, if you are interested in the position so we can schedule a meeting. Don't hesitate to reach out if you have any questions!