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LIVER Publication-derived

Modeling alcohol-associated liver disease in humans using adipose stromal or stem cell-derived organoids

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

👤 Guoyun Bi, Xuan Zhang, Weihong Li, Xin Lu, Xu He, Yaqiong Li, Rixing Bai, Haiyan Zhang ⏱ 13 days 📋 6 phases 🧫 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

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

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Bi et al., 2024. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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