Asymmetric Cellular Microenvironments

• Main Author: Rydholm, Susanna
• Format: Others
• Language: English
• Published: KTH, Cellens fysik 2008
• Subjects: Physics; Fysik
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4783; http://nbn-resolving.de/urn:isbn:978-91-7178-848-1

This thesis presents methods to combine 3D cell culture, microfluidics and gradients on a controlled cellular scale. 3D cultures in biological extracellular matrix gels or synthetic gels bridge the gap between organ-tissue cultures and traditional 2D cultures. A device for embedding, anchoring and culturing cells in a controlled 3D flow through micro-environment was designed and evaluated. The device was realized using an etched silicon pillar flow chamber filled with gel mixed with cells. The pillars anchor and stabilize the gel as well as increase the surface to volume ratio, permitting higher surface flow rates and improving diffusion properties. Within the structure cells were still viable and proliferating after six days of cultivation, showing that it is possible to perform medium- to-long term cultivation of cells in a controlled 3D environment. This concept was further developed to include controllable and time stable 3D microgradient environments. In this system stable diffusion gradients can be generated by the application of two parallel fluid flows with different composition against opposite sides of a gel plug with embedded cells. Culture for up to two weeks was performed showing cells still viable and proliferating. The cell tracer dye calcein was used to verify gradient formation as the fluorescent intensity in exposed cells was proportional to the position in the chamber. Cellular response to an applied stimulus was demonstrated by use of an adenosine triphosphate gradient where the onset of an intracellular calcium release also depends on cell position. === QC 20101119