Improving the generation of hepatocyte-like cells from pluripotent stem cells via high-efficient differentiation to definitive endoderm

doi:10.16352/j.issn.1001-6325.2023.07.1045

Abstract

Abstract: Objective To optimize the method of generating hepatocyte-like cells (HLCs) from pluripotent stem cells (PSCs)in vitro. Methods Considering the development of hepatocyte in vivo, a more efficient method was developed by optimizing biochemical factors to the stepwise differentiate PSCs into HLCs. Then the differentiation experiments were performed with human embryonic stem cell (hESC)line H9. The efficiency of the generation of HLCs was evaluated by morphological observation at different stages of differentiation. RT-qPCR and flow cytometry assay were used to examine the marker genes expression. Results A method of high-efficient hESCs differentiation to definitive endoderm (DE) was developed by adding cytokines activin A, bone morphogenetic protein 4(BMP4)and fibroblast growth factor 2(FGF2). It highly improved mRNA expression of marker genes at different stages. Venus fluorescence tracer and the results of flow-cytometry also showed that the population proportion of HLCs was significantly increased. Conclusions The high-efficient differentiation to DE improves differentiation efficiency of PSCs into HLCs.

Key words: differentiation, pluripotent stem cells, definitive endoderm, hepatocyte-like cell

CLC Number:

YAN Yucong, YANG Shuchun, JIA Yuyan, HUANG Yue. Improving the generation of hepatocyte-like cells from pluripotent stem cells via high-efficient differentiation to definitive endoderm[J]. Basic & Clinical Medicine, 2023, 43(7): 1045-1052.

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URL: http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/10.16352/j.issn.1001-6325.2023.07.1045

http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/Y2023/V43/I7/1045

References

[1] Satriano L, Lewinska M, Rodrigues PM, et al. Metabolic rearrangements in primary liver cancers: cause and consequences[J]. Nat Rev Gastroenterol Hepatol, 2019, 16: 748-766.
[2] Sia D, Villanueva A, Friedman SL, et al. Liver cancer cell of origin, molecular class, and effects on patient prognosis[J]. Gastroenterology, 2017, 152: 745-761.
[3] Forner A, Reig M, Bruix J.Hepatocellular carcinoma[J]. Lancet, 2018, 391: 1301-1314.
[4] Luce E, Messina A, Duclos Vallee JC, et al. Advanced techniques and awaited clinical applications for human pluripotent stem cell differentiation into hepatocytes[J]. Hepatology, 2021, 74: 1101-1116.
[5] Mun SJ.Generation of expandable human pluripotent stem cell-derived hepatocyte-like liver organoids[J]. J Hepatol, 2019, 71: 970-985.
[6] Si Tayeb K.Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells[J]. Hepatology, 2010, 51: 297-305.
[7] Thompson WL, Takebe T.Human liver model systems in a dish[J]. Dev Growth Differ, 2021, 63: 47-58.
[8] Touboul T.Generation of functional hepatocytes from human embryonic stem cells under chemically defined conditions that recapitulate liver development[J]. Hepatology, 2010, 51: 1754-1765.
[9] Xinaris C, Brizi V, Remuzzi G.Organoid models and applications in biomedical research[J]. Nephron, 2015, 130: 191-199.
[10] Ninomiya H, Mizwko K, Terada R, et al. Improved efficiency of definitive endoderm induction from human induced pluripotent stem cells in feeder and serum-free culture system in vitro[J].Cell Dev Biol Anim, 2015,51:1-8.
[11] Loh KM,Ang LT, Zhang J, et al. Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations[J]. Cell Stem Cell, 2014,14:237-252.
[12] Li S.Genetic and chemical screenings identify HDAC3 as a key regulator in hepatic differentiation of human pluripotent stem cells[J]. Stem Cell Reports, 2018, 11: 22-31.
[13] Akbari S, Arslan N, Senturk S, et al. Next-generation liver medicine using organoid models[J]. Front Cell Dev Biol, 2019, 7: 345, doi: 10.3389/fcell.2019.00345.
[14] Kretzschmar K, Clevers H.Organoids: modeling development and the stem cell niche in a dish[J]. Dev Cell, 2016, 38:590-600.
[15] Fatehullah A, Tan SH, Barker N.Organoids as an in vitro model of human development and disease[J]. Nat Cell Biol, 2016, 18:246-254.
[16] Li M, Izpisua Belmonte JC.Organoids-preclinical models of human disease[J]. N Engl J Med, 2019, 380: 569-579.