In this overview, we describe the Invasin methodology: what it can be used for, how it compares to existing organoid culturing technologies, and its main advantages and limitations.
What is the name of the methodology?
Invasin: Bacterial-expressed Invasin, an integrin-activating Yersinia protein, sustains long-term expansion of primary epithelial cells as 2D organoid sheets.
Developed by the Organoid Group.
What is the methodology good for?
We recently developed a novel organoid 2D sheet platform based on the coating of the bacterial integrin-activating protein of the Enteropathogenic Yersinia bacteria (Invasin).
Epithelial cells of different origins grow on Invasin in 2D and can be expanded and passaged long-term. In addition, polarization, junction formation and generation of various cell types were stable over time. Invasin, therefore, promises to represent a fully defined, versatile, cheap and animal-free alternative for BME/Matrigel.
What is/are the main advantages of this methodology over related technologies?
This 2D culturing approach holds practical advantages over the 3D “in gel” format: basal as well as apical domains are directly accessible for infection, uptake, or transport studies, while functional integrity of junctional barriers can readily be assessed. In addition, image analyses and high-throughput (drug) screening approaches are much simpler when executed in 2D.
What are the most important limitations of the methodology?
Not applicable.
What type of samples are compatible with methodology (yes, no, possibly)?
Cancer cell lines | Primary cells in culture | Organoids | Primary tissue |
Not Relevant | Not Relevant | Yes | Yes |
What future develops to the methodology are you planning, in any?
Coupling of Invasin to a gel for culturing organoids in 3D.
The generation and usage of image analyses and high-throughput (drug) screening protocols.
If someone outside your lab wants to use the methodology, what is the best option?
Protocols have been published and are continuously optimized. Purified Invasin protein can be obtained upon the signing of an MTA. Experience in how to culture organoids is essential.
Name one or more people in your lab that are experienced with the methodology
Joost Wijnakker and several other members of the Organoid Group.
Who originally developed the methodology?
The usage of Invasin as an alternative for BME/Matrigel for the growth of 2D organoids was developed by Wim de Lau and Joost Wijnakker.
The method has been published: Joost Wijnakker et al: doi: 10.1073/pnas.2420595121.
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