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LIVER Publication-derived

Alcohol-Related Liver Disease Organoids with Stromal Cells

Full SOP with Academic or a plan

Source Bi et al., 2024 · Department of Cell Biology, School of Basic Medical Science, Capital Medical University, Beijing, China · 10.1016/j.crmeth.2024.100778

Authors: Guoyun Bi, Xuan Zhang, Weihong Li, Xin Lu, Xu He, Yaqiong Li, Rixing Bai, Haiyan Zhang Duration: 13 days 6 phases Cell source: Human adipose stromal/stem cells (hASC)-derived hepatocyte-like cells (hHLCs) and endodermal progenitor cells (hEPCs)

Abstract

This protocol describes the generation of two types of hepatic organoids from human adipose stromal/stem cells (hASCs): hepatocellular organoids (hAHOs) composed primarily of hepatocyte-like cells, and more complex liver organoids (hALOs) containing hepatocytes and nonparenchymal cells. When exposed to ethanol, these organoids recapitulate steatosis, inflammation, and fibrosis characteristic of alcohol-associated liver disease (ALD), providing an in vitro human model for ALD pathogenesis research and therapeutic development.

Cell source
Human adipose stromal/stem cells (hASC)-derived hepatocyte-like cells (hHLCs) and endodermal progenitor cells (hEPCs)
Application
Disease modeling; alcohol-associated liver disease modeling

Protocol overview

17 steps across 6 phases

Phase 1 · hASC differentiation into hepatocyte-like cells (hHLCs) Day 0
  1. a Culture and differentiate hASCs into HLCs
Phase 2 · hAHO generation - cell seeding and spheroid formation Day 1-3
  1. a Harvest and trypsinize hHLCs
  2. b Wash and seed cells into ultra-low attachment plates
  3. c Centrifuge and incubate for spheroid formation
Phase 3 · hAHO generation - Matrigel embedding and expansion Day 3-13
  1. a Prepare Matrigel and expansion medium
  2. b Maintain hAHO culture with medium refreshment
  3. c Harvest organoids for analysis
Phase 4 · hALO generation - hEPC differentiation and spheroid formation Day 1-5
  1. a Culture and differentiate hASCs into endodermal progenitor cells (hEPCs)
  2. b Harvest and trypsinize hEPCs
  3. c Wash and seed hEPCs with retinoic acid
  4. d Centrifuge and incubate with RA for 5 days
Phase 5 · hALO generation - hepatocyte maturation phase Day 5-10
  1. a Switch to hepatocyte maturation medium
  2. b Culture for hepatic maturation with medium refreshment
  3. c Harvest hALOs for analysis
Phase 6 · Alcohol-associated liver disease (ALD) modeling - ethanol treatment 3-day treatment
  1. a Prepare ethanol treatment medium
  2. b Incubate organoids with ethanol
  3. c Harvest ethanol-treated organoids

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Attribution

This SOP was written by Organthis based on the published method in Bi et al., 2024. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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