Unraveling the beneficial effects of 3D microenvironments:
how do stem cell organoids enhance chondrogenesis?
Native tissues are naturally three dimensional. However, traditional cell culture is performed on two dimensional monolayers. In recent years, we have developed microfabrication techniques that allow us to form organoids (micrometer sized tissues) in a high throughput manner. These organoids have consistently been demonstrated to outperform traditional cultures in terms of tissue formation, both in vitro and in vivo.
In this project, we test our hypothesis why three dimensional stem cell cultures outperform two dimensional cultures. This will be based on exploiting stem fluorescent microscopy and biochemical analysis stem cell organoids. Validating our hypothesis will strongly contribute to our understanding of (stem) cell biology and allow us to intelligently design novel approaches to improve engineered tissues using this unique mechanism.
Specifically, you will be involved in the microfabrication of biomaterial microwell arrays, perform stem cell cultures, fluorescent confocal microscopy, biochemical analysis, and study the effects of preselected inhibitors on stem cell fate. This project will therefore provide aspiring students with an interdisciplinary skillset and knowledge framework on stem cells, microfabrication, biomaterials, and tissue engineering.
Figure 1: Pilot experiments illustrating (A) formation of a stem cell organoid over time. (B) This unique culture system induces the expression chondrogenic master regulators such as SOX9, which leads to (C) higher expression of cartilage related genes and (D) potently enhanced formation of cartilage. We have developed a model that has generated a hypothesis why this 3D microenvironment stimulates stem cell differentiation, which will be validated in this project.