3D Organoids from primary melanoma cell lines and from iPSc-derived neural crest stem cells
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- 3D culture
- single-cell RNA seq
- single-cell ATAC seq
We propose to generate three-dimensional tumoroids from the primary melanoma cell lines, as well as 3D organoids from the iPSc-derived neural crest stem cells. We will use the AggreWell system (STEMCELL Technologies) to generate uniform, size-controlled three-dimensional spheroids. After 5 days in the AggreWell plate, the spheroids are moved to a PEG-based artificial ECM hydrogel (Gjorevski et al.; Nature Protocols 2017). The organoids can be cultured for weeks in these PEG-droplets. At different time points during organoid culture, organoids will be used for immunostaining and/or for single-cell sequencing. We will dissociate the PEG gel to obtain single cells by use of the cell-dissociation enzyme TrypLE.
- Biosafety cabinet ;
- Cell incubator CO2-connected ;
- Centrifuge for plates.
- Published in peer reviewed journal
Pros, cons & Future potential
3D organoids mimic tissue architecture heterogenous cell culture to study cellular differentiation enhancer testing.
Not every cell type/tissue can be studied.
Different cell types are studied to form organoids.
Drug application: concentration and activity can be tested.
References, associated documents and other information
Gjorevski N & Lutolf MP. Synthesis and characterization of well-defined hydrogel matrices and their application to intestinal stem cell and organoid culture. Nature Protocols 12 (11); 2263-2274 (2017)
Contact personValerie Christiaens
OrganisationsVIB - KU Leuven
Department of Brain and Disease Rearch
Lab of Computational Biology