Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens

Bastien Laperrousaz; Stéphanie Porte; Sophie Gerbaud; Ville Härmä; Frederique Kermarrec; Virginie Hourtané; Frédéric Bottausci; Xavier Gidrol; Nathalie Picollet-D’hahan

Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens

Bastien Laperrousaz ¹ Stéphanie Porte ¹ Sophie Gerbaud ² Ville Härmä ¹ Frederique Kermarrec ² Virginie Hourtané ³ Frédéric Bottausci ³ Xavier Gidrol ^{1, *} Nathalie Picollet-D’hahan ^{2, *}

* Corresponding author

1 BGE - UMR S1038 - Laboratoire de Biologie à Grande Échelle

2 BIOMICS - Biomicrotechnologie et génomique fonctionnelle

BGE - UMR S1038 - Laboratoire de Biologie à Grande Échelle, UGA - Université Grenoble Alpes

3 CEA-LETI - Laboratoire d'Electronique et des Technologies de l'Information

Abstract : Organoid cultures in 3D matrices are relevant models to mimic the complex in vivo environment that supports cell physiological and pathological behaviors. For instance, 3D epithelial organoids recapitulate numerous features of glandular tissues including the development of fully differentiated acini that maintain apico-basal polarity with hollow lumen. Effective genetic engineering in organoids would bring new insights in organogenesis and carcinogenesis. However, direct 3D transfection on already formed organoids remains challenging. One limitation is that organoids are embedded in extracellular matrix and grow into compact structures that hinder transfection using traditional techniques. To address this issue, we developed an innovative approach for transgene expression in 3D organoids by combining single-cell encapsulation in Matrigel microbeads using a microfluidic device and electroporation. We demonstrate that direct electroporation of encapsulated organoids reaches up to 80% of transfection efficiency. Using this technique and a morphological read-out that recapitulate the different stages of tumor development, we further validate the role of p63 and PTEN as key genes in acinar development in breast and prostate tissues. We believe that the combination of controlled organoid generation and efficient 3D transfection developed here opens new perspectives for flow-based high-throughput genetic screening and functional genomic applications.

Document type :

Journal articles

Domain :

Engineering Sciences [physics]

Complete list of metadatas

https://hal-cea.archives-ouvertes.fr/cea-02183666
Contributor : Marianne Leriche <>
Submitted on : Monday, July 15, 2019 - 3:12:35 PM
Last modification on : Wednesday, July 17, 2019 - 1:04:45 AM

Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens

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Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens

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