STAR Protocols (Jun 2023)

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

Journal volume & issue
Vol. 4, no. 2
p. 102333

Abstract

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Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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