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

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.

Validating Well-Functioning Hepatic Organoids for Toxicity Evaluation

“…Despite its vital role in drug detoxification, current in vitro and in vivo methods fall short of fully replicating human liver physiology owing to chromosomal abnormalities, functional immaturity, and interspecies variations [16]. To address this gap, we established a liver organoid model that mimicked the functional and gene expression aspects of the human liver.…”

“…Liver similarity analysis was conducted as previously described [16,17]. The liverspecific gene expression panel (LiGEP) algorithm was quantified using the fragments per kilobase of transcript per million mapped reads (FPKM) value.…”