citations

                    Standard operating protocol of hepatic organoid 
differentiation from human induced pluripotent stem 
cells 
                

Authors:

Min Jung Kim, Jaeseo Lee , Seon Ju Mun, Myung Jin Son, Jung-Hyun Kim

In:

Source: Other
Publication Date: ()
Issue: 2: e5

Research Area:

Stem Cells

Toxicology

Drug Discovery

Cells used in publication:

Induced Pluripotent Stem Cell (iPS), human: Species: human; Tissue Origin:

Culture Media:

Endothelial Cell Growth Medium 2

Hepatocyte Culture Medium

Experiment

3) Stage 3: mature hepatocytes To induce differentiation to mature hepatocytes in a 2D culture system, the medium was replaced with Hepatocyte Culture Medium (CC-3198; Lonza, Basel, Swiss) without epidermal growth factor, mixed with Endothelial Cell Growth Medium-2 (CC-3162; Lonza, Basel, Swiss) (1:1), adjusted to a 2.5% fetal bovine serum concentration with 100 nM dexamethasone (D4902; Sigma-Aldrich, St. Louis, MO, USA), 20 ng/mL OSM (295-OM-050; R&D Systems, Minneapolis, MN, USA), and 10 ng/mL human hepatocyte growth factor (100-39; Peprotech) for 4 days and incubated in 5% hypoxia. The cells were then further cultured in normoxia for 7 days.

Abstract

Mature liver organoids are promising cell sources for research to understand the pathology underlying a variety of conditions affecting the liver, including end-stage chronic liver disease. Although several methods exist for the differentiation of mature hepatic organoids derived from human induced pluripotent stem cells (hiPSCs), organoid generation can fail due to various experimental culture conditions. Therefore, we established a standard operating protocol for generating mature and expandable hepatic organoids derived from hiPSCs, and we made the starting materials available to facilitate the wide use of the protocol.

Open in PubMed

Standard operating protocol of hepatic organoid differentiation from human induced pluripotent stem cells