한빛사논문
- 조회579
Soraya Salas-Silva a,b,1, Yohan Kim a,b,c,d,1, Tae Hun Kim a,b, Myounghoi Kim a,b, Daekwan Seo e, Jeonghoon Choi e, Valentina M. Factor f, Haeng Ran Seo g, Yeonhwa Song g, Gyu Sung Choi h, Yun Kyung Jung a, Kungsik Kim a, Kyeong Geun Lee a, Jaemin Jeong i, Ji Hyun Shin a,b, Dongho Choi a,b,j
aDepartment of Surgery, Hanyang University College of Medicine, Seoul, 04763, Republic of Korea
bResearch Institute of Regenerative Medicine and Stem Cells, Hanyang University, Seoul, 04763, Republic of Korea
cMax Planck Institute of Molecular Cell Biology and Genetics, 01307, Dresden, Germany
dDepartment of MetaBioHealth, Sungkyunkwan University, Suwon, 16419, Republic of Korea
eOffice of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Researcj, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA
fLaboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA
gAdvanced Biomedical Research Laboratory, Institute Pasteur Korea, 16, Daewangpangyo-ro 712-beon gil, Bundang-gu, Seongnam-si, Gyeonggi-do, 13488, Republic of Korea
hDepartment of General Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea
iLaboratory of Radiation Exposure and Therapeutics, National Radiation Emergency Medical Center, KIRAMS, Republic of Korea
jDepartment of HY-KIST Bio-convergence, Hanyang University, Seoul, 04763, Republic of Korea
1These authors contributed equally to this study as co-first authors.
Corresponding authors: Ji Hyun Shin, Dongho Choi
Abstract
Background & aims
Several types of human stem cells from embryonic (ESCs) and induced pluripotent (iPSCs) to adult tissue-specific stem cells are commonly used to generate 3D liver organoids for modeling tissue physiology and disease. We have recently established a protocol for direct conversion of primary human hepatocytes (hPHs) from healthy donor livers into bipotent progenitor cells (hCdHs). Here we extended this culture system to generate hCdH-derived liver organoids for diverse biomedical applications.
Methods
To obtain hCdHs, hPHs were cultured in reprogramming medium containing A83-01 and CHIR99021 for 7 days. Liver organoids were established from hCdHs (hCdHOs) and human liver cells (hLOs) using the same donor livers for direct comparison, as well as from hiPSCs. Organoid properties were analyzed by standard in vitro assays. Molecular changes were determined by RT-qPCR and RNA-seq. Clinical relevance was evaluated by transplantation into FRG mice, modeling of alcohol-related liver disease (ARLD) and in vitro drug-toxicity tests.
Results
hCdHs were clonally expanded as organoid cultures with low variability between starting hCdH lines. Similar to the hLOs, hCdHOs stably maintained stem cell phenotype based on accepted criteria. However, hCdHOs had an advantage over hLOs in terms of EpCAM expression, efficiency of organoid generation and capacity for directed hepatic differentiation as judged by molecular profiling, albumin secretion, glycogen accumulation, and CYP450 activities. Accordingly, FRG mice transplanted with hCdHOs survived longer than mice injected with hLOs. When exposed to ethanol, hCdHOs developed stronger ALD phenotype than hLOs as evidenced by transcriptional profiling, lipid accumulation and mitochondrial dysfunction. In drug-induced injury assays in vitro, hCdHOs showed a similar or higher sensitivity response than hPHs.
Conclusion
hCdHOs provide a novel patient-specific stem cell-based platform for regenerative medicine, toxicology testing and modeling liver diseases.
논문정보
- Research article
- 2023년 10월 (BRIC 등록일 2023-10-25)
- 국내(교신)+국외 연구진
- 생명과학 > 세포생물학
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