Designer matrices for intestinal stem cell and organoid culture

Autor: Paloma Ordóñez-Morán, Maiia E. Bragina, Norman Sachs, Sonja Giger, Matthias P. Lutolf, Hans Clevers, Andrea Manfrin, Nikolce Gjorevski
Přispěvatelé: Hubrecht Institute for Developmental Biology and Stem Cell Research
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Cellular differentiation
Cell Culture Techniques
Nanotechnology
02 engineering and technology
Research Support
complex mixtures
Hydrogel
Polyethylene Glycol Dimethacrylate
Article
Extracellular matrix
Tissue Culture Techniques
03 medical and health sciences
Mice
Laminin
Organoid
Cell Adhesion
Journal Article
Animals
Humans
Cell Lineage
Stem Cell Niche
Cell adhesion
Non-U.S. Gov't
General
Cell Shape
Cell Proliferation
Medicine(all)
Multidisciplinary
biology
Stem Cells
Research Support
Non-U.S. Gov't
technology
industry
and agriculture
Cell Differentiation
021001 nanoscience & nanotechnology
Cell biology
Extracellular Matrix
Fibronectins
Fibronectin
Intestines
Organoids
030104 developmental biology
Proteolysis
biology.protein
Stem cell
0210 nano-technology
Function (biology)
Zdroj: Nature, 539(7630), 560-564. Nature Publishing Group
ISSN: 0028-0836
Popis: 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.
Databáze: OpenAIRE
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