Generation of liver mesenchyme and ductal cell organoid co-culture using cell self-aggregation and droplet microfluidics

Anna M Dowbaj; Timo N Kohler; Lucía Cordero-Espinoza; Florian Hollfelder; Meritxell Huch

doi:10.1016/j.xpro.2023.102333

Generation of liver mesenchyme and ductal cell organoid co-culture using cell self-aggregation and droplet microfluidics

STAR Protoc. 2023 Jun 2;4(2):102333. doi: 10.1016/j.xpro.2023.102333. Online ahead of print.

Authors

Anna M Dowbaj¹, Timo N Kohler², Lucía Cordero-Espinoza³, Florian Hollfelder⁴, Meritxell Huch⁵

Affiliations

¹ Max Planck Institute of Molecular Cell Biology and Genetics, Dresden 01307, Germany.
² Wellcome Trust-Medical Research Council Stem Cell Institute Cambridge, Cambridge CB2 1QR, UK; Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
³ Wellcome Trust-Medical Research Council Stem Cell Institute Cambridge, Cambridge CB2 1QR, UK; Wellcome Trust/Cancer Research UK Gurdon Institute, Cambridge CB2 1QN, UK; Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK.
⁴ Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
⁵ Max Planck Institute of Molecular Cell Biology and Genetics, Dresden 01307, Germany; Wellcome Trust-Medical Research Council Stem Cell Institute Cambridge, Cambridge CB2 1QR, UK. Electronic address: [email protected].

Abstract

Within the peri-portal region of the adult liver, portal fibroblasts exist in close proximity to epithelial ductal/cholangiocyte cells. However, the cellular interactions between them are poorly understood. Here, we provide two co-culture techniques to incorporate liver portal mesenchyme into ductal cell organoids, which recapitulate aspects of their cellular interactions in vitro. We integrate several techniques from mesenchyme isolation and expansion to co-culture by microfluidic cell co-encapsulation or 2D-Matrigel layer. The protocol is easily adaptable to other cells from other organs. For complete information on the generation and use of this protocol, please refer to Cordero-Espinoza et al.¹.

Keywords: Cell Biology; Organoids; Stem Cells.