Protocol on mapping spatial organization of mammalian cells published by Wisniewski et al.
Tissue Repair PhD student Darren Wisniewski et al publish method to reveal spatial organization within mammalian cultures.
Darren Wisniewski, third-year Wellcome Trust Tissue Repair PhD student in the Lowell lab at the the MRC Centre for Regenerative Medicine, has reported a method using micropatterning together with quantitative imaging to reveal spatial organization within mammalian cultures. The technique is easy to establish in a standard cell biology laboratory and offers a tractable system to study patterning in vitro.
Darren and co-author Dr Guillaume Blin explain and show their method in the JoVE video that accompanies the paper.
A fundamental goal in biology is to understand how patterns emerge during development. Several groups have shown that patterning can be achieved in vitro when stem cells are spatially confined onto micropatterns, thus setting up experimental models which offer unique opportunities to identify, in vitro, the fundamental principles of biological organisation.
Here we describe our own implementation of the methodology. We adapted a photo-patterning technique to reduce the need for specialized equipment to make it easier to establish the method in a standard cell biology laboratory. We also developed a free, open-source and easy to install image analysis framework in order to precisely measure the preferential positioning of sub-populations of cells within colonies of standard shapes and sizes. This method makes it possible to reveal the existence of patterning events even in seemingly disorganized populations of cells. The technique provides quantitative insights and can be used to decouple influences of the environment (e.g., physical cues or endogenous signaling), on a given patterning process.