Designer matrices for intestinal stem cell and organoid culture

By N. Gjorevski, Norman Sachs, Andrea Manfrin, Sonja Giger, Maiia E. Bragina, P. Ordóñez-Morán, Hans Clevers, M. Lutolf. Nature, 2016

1,280 citations recorded by the source corpus.

Epithelial organoids recapitulate multiple aspects of real organs, making them promising models of organ development, function and disease. However, the full potential of organoids in research and therapy has remained unrealized, owing to the poorly defined animal-derived matrices in which they are grown. Here we used modular synthetic hydrogel networks to define the key extracellular matrix (ECM) parameters that govern intestinal stem cell (ISC) expansion and organoid formation, and show that separate stages of the process require different mechanical environments and ECM components. In particular, fibronectin-based adhesion was sufficient for ISC survival and proliferation. High matrix stiffness significantly enhanced ISC expansion through a yes-associated protein 1 (YAP)-dependent mechanism. ISC differentiation and organoid formation, on the other hand, required a soft matrix and laminin-based adhesion. We used these insights to build a fully defined culture system for the expansion of mouse and human ISCs. We also produced mechanically dynamic matrices that were initially optimal for ISC expansion and subsequently permissive to differentiation and intestinal organoid formation, thus creating well-defined alternatives to animal-derived matrices for the culture of mouse and human stem-cell-derived organoids. Our approach overcomes multiple limitations of current organoid cultures and greatly expands their applicability in basic and clinical research. The principles presented here can be extended to identify designer matrices that are optimal for long-term culture of other types of stem cells and organoids.

Subfields: Biomedical Engineering, Cell Biology, Oncology

View Paper: https://www.semanticscholar.org/paper/88e4d5585e5b4bc8ae3d77833139f24d64a3dae0

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Biomedical Engineering

1. S. Piccolo
2. Hans Clevers

In Cell Biology

1. Guo-li Ming
2. R. Nusse
3. Hongjun Song
4. S. Piccolo
5. Hans Clevers
6. E. Voest
7. K. Guan

In Oncology

1. R. Nusse
2. Guo-li Ming
3. Hongjun Song
4. Hans Clevers
5. Ton N. Schumacher
6. S. Piccolo
7. Pengfei Lu

Citations in the corpus

• Wnt/ b -Catenin Signaling, Disease, and Emerging Therapeutic Modalities (2017, background): “Another technical improvement involves the replacement of Matrigel by a synthetic hydrogel (Gjorevski et al., 2016).”
• Brain organoids: advances, applications and challenges (2019, background): “The use of variable ingredients, such as animal-derived ECM (Matrigel), in culture protocols should also be minimized, and recombinant growth factors should be replaced by small molecules whenever applicable (Cruz-Acuña et al., 2017; Gjorevski et al., 2016).”
• A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity. (2019, background): “The use of a fully defined culture medium enhances the reproducibility of cultures (Gjorevski et al., 2016) and facilitates its use in future clinical applications.”
• YAP/TAZ-Dependent Reprogramming of Colonic Epithelium Links ECM Remodeling to Tissue Regeneration (2018, background): “Recent evidence with artificially designed matrices indicates that conditions that sustain YAPactivation canmaintain primary epithelial cells in vitro (Gjorevski et al., 2016).”
• Generation of tumor-reactive T cells by co-culture of peripheral blood lymphocytes and tumor organoids (2018, positive): “Of note, strategies to expand organoids in synthetic matrices have been described recently (Gjorevski et al., 2016) and could serve as alternatives to bypass reactivity to animal antigen.”