한빛사논문
- 조회1,489
Gyunggyu Leea,1, Hyemin Kimb,1, Ji Young Parka, Gyeongmin Kima, Jiyou Hanc, Seok Chungd,e, Ji Hun Yange, Jang Su Jeonf, Dong-Hun Woog, Choongseong Hang, Sang Kyum Kimf,*, Han-Jin Parkb,*, Jong-Hoon Kima,*
aLaboratory of Stem Cells and Tissue Regeneration, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea
bDepartment of Predictive Toxicology, Korea Institute of Toxicology, Daejeon 34114, Korea
cDepartment of Biological Sciences, Hyupsung University, Hwasung-si 18330, Korea
dSchool of Mechanical Engineering, Korea University, Seoul 20841, Korea
eKU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Korea
fChungnam National University, Daejeon 34134, Korea
gLaboratory of Stem Cells, NEXEL Co., Ltd., Seoul 02580, Korea
1These authors contributed equally to this work.
*Corresponding author.
Abstract
Recent advances in pluripotent stem cell technology provide an alternative source of human hepatocytes to overcome the limitations of current toxicity tests. However, this approach requires optimization and standardization before it can be used as a fast and reliable toxicity screening system. Here, we designed and tested microwell culture platforms with various diameters. We found that large quantities of uniformly-sized hepatocyte-like cell (HLC) spheroids (3D-uniHLC-Ss) could be efficiently and reproducibly generated in a short period time from a small number of differentiating human pluripotent stem cells (hPSCs). The hPSC-3D-uniHLC-Ss that were produced in 500-μm diameter microwells consistently exhibited high expressions of hepatic marker genes and had no significant signs of cell death. Importantly, a hepatic master gene hepatocyte nuclear factor 4α (HNF4α) was maintained at high levels, and the epithelial-mesenchymal transition was significantly attenuated in hPSC-3D-uniHLC-Ss. Additionally, when compared with 3D-HLC-Ss that were produced in other 3D platforms, hPSC-3D-uniHLC-Ss showed significantly higher hepatic gene expressions and drug-metabolizing activity of the enzyme, CYP3A4. Imaging-based drug toxicity studies demonstrated that hPSC-3D-uniHLC-Ss exhibited enhanced sensitivity to various hepatotoxicants, compared to HLCs, which were differentiated under 2D conditions. Precise prediction of drug-induced hepatotoxicity is a crucial step in the early phases of drug discovery. Thus, the hPSC-3D-uniHLC-Ss produced using our microwell platform could be used as an imaging-based toxicity screening system to predict drug hepatotoxicity.
Keywords : stem cell, spheroid, microwell, hepatocyte, toxicity
논문정보
- Research article
- 2020년 11월 (BRIC 등록일 2020-11-24)
- 국내 연구진
- 생명과학 > 세포생물학
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