Protocol for automated production of human stem cell derived liver spheres

STAR Protoc. 2021 Apr 30;2(2):100502. doi: 10.1016/j.xpro.2021.100502. eCollection 2021 Jun 18.

Abstract

This protocol describes how to produce human liver spheres from pluripotent stem cell-derived hepatic progenitors, endothelial cells, and hepatic stellate cells. Liver spheres form by self-assembly in microwells, generating up to 73 spheres per well of a 96-well plate. This process was automated using liquid handling and pipetting systems, permitting cost-effective scale-up and reducing sphere variability. In its current form, this system provides a powerful tool to generate human liver tissue for disease modeling and drug screening. For complete details on the use and execution of this protocol, please refer to Lucendo-Villarin et al. (2020) (https://doi.org/10.1088/1758-5090/abbdb2).

Keywords: Cell Differentiation; Metabolism; Stem Cells; Tissue Engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation, Laboratory*
  • Cell Culture Techniques*
  • Cell Differentiation*
  • Humans
  • Liver* / cytology
  • Liver* / metabolism
  • Pluripotent Stem Cells* / cytology
  • Pluripotent Stem Cells* / metabolism
  • Spheroids, Cellular* / cytology
  • Spheroids, Cellular* / metabolism