Culture and analysis of kidney tubuloids and perfused tubuloid cells-on-a-chip

Linda Gijzen, Fjodor A Yousef Yengej, Frans Schutgens, Marianne K Vormann, Carola M E Ammerlaan, Arnaud Nicolas, Dorota Kurek, Paul Vulto, Maarten B Rookmaaker, Henriette L Lanz, Marianne C Verhaar, Hans Clevers

Research output: Contribution to journal/periodicalArticleScientificpeer-review

34 Citations (Scopus)


Advanced in vitro kidney models are of great importance to the study of renal physiology and disease. Kidney tubuloids can be established from primary cells derived from adult kidney tissue or urine. Tubuloids are three-dimensional multicellular structures that recapitulate tubular function and have been used to study infectious, malignant, metabolic, and genetic diseases. For tubuloids to more closely represent the in vivo kidney, they can be integrated into an organ-on-a-chip system that has a more physiological tubular architecture and allows flow and interaction with vasculature or epithelial and mesenchymal cells from other organs. Here, we describe a detailed protocol for establishing tubuloid cultures from tissue and urine (1-3 weeks), as well as for generating and characterizing tubuloid cell-derived three-dimensional tubular structures in a perfused microfluidic multi-chip platform (7 d). The combination of the two systems yields a powerful in vitro tool that better recapitulates the complexity of the kidney tubule with donor-specific properties.

Original languageEnglish
Pages (from-to)2023-2050
Number of pages28
JournalNature Protocols
Issue number4
Publication statusPublished - Apr 2021


  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Cell Fractionation
  • Child
  • Child, Preschool
  • Electric Impedance
  • Female
  • Fluorescent Dyes/chemistry
  • Humans
  • Infant
  • Kidney Tubules/growth & development
  • Lab-On-A-Chip Devices
  • Male
  • Membrane Transport Proteins/metabolism
  • Microfluidics
  • Middle Aged
  • Organoids/growth & development
  • Perfusion
  • Rats
  • Tissue Culture Techniques/methods
  • Young Adult


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