Temporal analyses of postnatal liver development and maturation by single-cell transcriptomics
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Abstract
The postnatal development and maturation of the liver, the major metabolic organ, are inadequately understood. We have analyzed 52,834 single-cell transcriptomes and identified 31 cell types or states in mouse livers at postnatal days 1, 3, 7, 21, and 56. We observe unexpectedly high levels of hepatocyte heterogeneity in the developing liver and the progressive construction of the zonated metabolic functions from pericentral to periportal hepatocytes, which is orchestrated with the development of sinusoid endothelial, stellate, and Kupffer cells. Trajectory and gene regulatory analyses capture 36 transcription factors, including a circadian regulator, Bhlhe40, in programming liver development. Remarkably, we identified a special group of macrophages enriched at day 7 with a hybrid phenotype of macrophages and endothelial cells, which may regulate sinusoidal construction and Treg-cell function. This study provides a comprehensive atlas that covers all hepatic cell types and is instrumental for further dissection of liver development, metabolism, and disease.
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Numerous changes in mRNA abundance and splicing occur within the first 3 weeks of life, as hematopoiesis and proliferative activity are reduced, and gene products involved in metabolism are induced.
“Numerous changes in mRNA abundance and splicing occur within the first 3 weeks of life (Bhate et al. 2015; Chembazhi et al. 2021; Liang et al. 2022), as hematopoiesis and proliferative activity are reduced, and gene products involved in metabolism are induced (Zaret 2002; Gordillo et al. 2015).”
From ESRP2–microRNA-122 axis promotes the postnatal onset of liver polyploidization and maturation (2025) · cites this paper as Liang et al. 2022
The Results sections on hepatocyte subpopulations and transcription factors report changes in gene expression from day 1 through day 21, supporting the mRNA-abundance part of the cited clause. They do not report changes in splicing over those three weeks; the mention of a spliced mRNA describes one transcript, not a temporal change. The later claims about hematopoiesis, proliferation, and metabolism are attached to different citations in the citing sentence. Page numbers are not available in our copy. (LLM, conf=0.88)
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Gut–liver axis: barriers and functional circuits
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Scalable production of tissue-like vascularized liver organoids from human PSCs
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OmicVerse: a framework for bridging and deepening insights across bulk and single-cell sequencing
Zeng, Z., Ma, Y., Hu, L., Tan, B., Liu, P., Wang, Y., Xing, C., Xiong, Y., & Du, H. (2024) · Nature Communications · doi:10.1038/s41467-024-50194-3 · Cited by 102 · see citing works
Macrophage heterogeneity in the single-cell era: facts and artifacts
Hume, D. A., Millard, S. M., & Pettit, A. R. (2023) · Blood · doi:10.1182/blood.2023020597 · Cited by 78 · see citing works
Human iPSC-liver organoid transplantation reduces fibrosis through immunomodulation
Tadokoro, T., Murata, S., Kato, M., Ueno, Y., Tsuchida, T., Okumura, A., Kuse, Y., Konno, T., Uchida, Y., Yamakawa, Y., et al. (2024) · Science Translational Medicine · doi:10.1126/scitranslmed.adg0338 · Cited by 58 · see citing works
Priming and Maintenance of Adaptive Immunity in the Liver
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Fetal liver development and implications for liver disease pathogenesis
Lotto, J., Stephan, T. L., & Hoodless, P. A. (2023) · Nature Reviews Gastroenterology & Hepatology · doi:10.1038/s41575-023-00775-2 · Cited by 47 · see citing works
Single-cell and spatially resolved transcriptomics for liver biology
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PGLYRP2 drives hepatocyte-intrinsic innate immunity by trapping and clearing hepatitis B virus
Li, Y., Ma, H., Zhang, Y., He, T., Li, B., Ren, H., Feng, J. F., Sheng, J., Li, K., Yu, Q., et al. (2025) · Journal of Clinical Investigation · doi:10.1172/jci188083 · Cited by 36 · see citing works
From multitude to singularity: An up-to-date overview of scRNA-seq data generation and analysis
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Contamination of isolated mouse Kupffer cells with liver sinusoidal endothelial cells
Hume, D. A., Offermanns, S., & Bonnavion, R. (2022) · Immunity · doi:10.1016/j.immuni.2022.06.010 · Cited by 31 · see citing works
Determination of key events in mouse hepatocyte maturation at the single-cell level
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Single-cell dynamics of liver development in postnatal pigs
Rao, L., Cai, L., & Huang, L. (2023) · 中国科学通报:英文版 · doi:10.1016/j.scib.2023.09.021 · Cited by 26 · see citing works
Kupffer cell reverse migration into the liver sinusoids mitigates neonatal sepsis and meningitis
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Kupffer cells in liver homeostasis and disease: from immune sentinels to metabolic gatekeepers
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Decoding liver fibrogenesis with single-cell technologies
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Developmental programming of tissue-resident macrophages
Viola, M. F., Taveras, E. F., & Mass, E. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1475369 · Cited by 20 · see citing works
Immune Cell Contribution to Mammary Gland Development
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Insights into the gut-liver axis: mechanisms and emerging therapies in hepatocellular carcinoma
Hu, Y., Gao, M., Chenghuang, J., & Bao, R. (2025) · Frontiers in Pharmacology · doi:10.3389/fphar.2025.1595853 · Cited by 16 · see citing works
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SHMT2 reduces fatty liver but is necessary for liver inflammation and fibrosis in mice
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The mouse multi-organ proteome from infancy to adulthood
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Nagy, D., Maude, H., Birdsey, G. M., RANDI, A. M., & Cebola, I. (2023) · Journal of Molecular Endocrinology · doi:10.1530/jme-23-0026 · Cited by 12 · see citing works
Deciphering the temporal transcriptional landscape of human fetal leptomeninges
Sun, L., Liu, P., Guo, J., Fang, C., Li, L., Liu, Y., Tan, Y., Zhang, W., Zhao, R., Zhang, F., et al. (2024) · Brain · doi:10.1093/brain/awae397 · Cited by 12 · see citing works
Myeloid cells in alcoholic liver diseases: Mechanism and prospect
Xu, W., Wu, M., Chen, B., & Wang, H. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.971346 · Cited by 11 · see citing works
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Unveiling the power of microenvironment in liver regeneration: an in-depth overview
Hu, Y., Wang, R., An, N. A., Li, C., Wang, Q., Cao, Y., Li, C., Liu, J., & Wang, Y. (2023) · Frontiers in Genetics · doi:10.3389/fgene.2023.1332190 · Cited by 12 · see citing works
Single-cell analysis of diquat-induced oxidative stress and its impact on organ-specific toxicity
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ADAMTS18-fibronectin interaction regulates the morphology of liver sinusoidal endothelial cells
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Jag1 insufficiency alters liver fibrosis via T cell and hepatocyte differentiation defects
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Kaneko, K., Liang, Y., Liu, Q., Zhang, S., Scheiter, A., Song, D., & Feng, G. (2023) · eLife · doi:10.7554/elife.86824.3 · Cited by 6 · see citing works
Self-citationTranscriptomic Profile of the Mouse Postnatal Liver Development by Single-Nucleus RNA Sequencing
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ESRP2–microRNA-122 axis promotes the postnatal onset of liver polyploidization and maturation
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Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs
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Proenkephalin produced by neonatal T-bet+ Treg cells promotes periportal hepatocyte maturation
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Self-citationHepatocyte differentiation requires anisotropic expansion of bile canaliculi
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IGF2 mediates Hippo signaling to control liver sizePreprint
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Aberrant differentiation and proliferation of hepatocytes in chronic liver injury and liver tumors
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Multifaceted Role of ILCs in Metabolic Adaptations: From Adult to Neonate
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UFMylation: biological mechanisms, functions, and clinical implications
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LDL-Bound PCSK9 Has a Slower Clearance Kinetic and Higher Use for HSPGs Than Free-PCSK9—Brief Report
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ESRP2-microRNA-122 axis directs the postnatal onset of liver polyploidization and maturationPreprint
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Spatiotemporally restricted Hippo signalings instruct the fate and maturation of hepatobiliary cellsPreprint
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MittoSAM-dependent lipoylation controls postnatal heart development via metabolic remodelingPreprint
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Hepatocyte differentiation requires anisotropic expansion of bile canaliculiPreprint
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Single-Cell Sequencing Analysis of the Differentiation of Mouse Hepatic Oval Cells
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Peroxisomes and their role in pediatric diseases
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The gut microbiome promotes detoxification responses to an environmental toxicantPreprint
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Postnatal hepatocyte proliferation – challenge and opportunity for gene therapy
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Single-cell activation screen identifies hepatic maturation regulators with zonal resolution
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Additional file 8 of Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs
Zhu, J., Guan, X., Yi, L.-L., Xu, H., Li, Q.-Y., Cheng, W.-J., Xie, Y., Li, W.-Z., Zhao, H., Wei, H.-J., et al. (2024) · Figshare · doi:10.6084/m9.figshare.26652858
Additional file 1 of Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs
Zhu, J., Guan, X., Lanlan, Y., Xu, H., Li, Q.-Y., Cheng, W.-J., Xie, Y., Li, W.-Z., Zhao, H., Wei, H.-J., et al. (2024) · Figshare · doi:10.6084/m9.figshare.26652837
Additional file 5 of Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs
Zhu, J., Guan, X., Yi, L.-L., Xu, H., Li, Q., Cheng, W.-J., Xie, Y., Li, W.-Z., Zhao, H.-Y., Wei, H.-J., et al. (2024) · Figshare · doi:10.6084/m9.figshare.26652849
- Authors
- Liang, Y., Kaneko, K., Xin, B., Lee, J., Sun, X., Zhang, K., & Feng, G.-S.
- Affiliations
- University of California San Diego (US)
- Corresponding author
- Feng, Gen-Sheng
- Venue
- Developmental Cell
- Publisher
- Elsevier BV
- Published
- 2022-02
- Volume
- 57
- Issue
- 3
- Pages
- 398-414.e5
- ISSN
- 1534-5807 (ISSN-L)
- PMC
- PMC8842999
- OpenAlex
- W4221105933
- Cited by
- 116
- References
- 88
- Citations by year
- 2026: 28 · 2025: 29 · 2024: 27 · 2023: 25 · 2022: 7
- FWCI
- 8.25
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 371 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Developmental Cell (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- American Association for the Study of Liver Diseases
- National Institutes of Health (R01DK128320, R01CA236074, R01CA239629, S10 OD026929)
- National Institute of Neurological Disorders and Stroke (P30NS047101)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- postnatal liver development, liver maturation, hepatocyte heterogeneity, zonation, sinusoidal endothelial cells, hepatic stellate cells, Kupffer cells, hepatic macrophages, BHLHE40, circadian regulation, transcription factors, regulatory networks, single-cell transcriptomics, mouse liver, regulatory T cells
- MeSH terms
- RNA-Seq, Animals, Animals, Newborn, Cell Communication, Hematopoiesis, Liver, Macrophages, Mesoderm, Time Factors, Transcription Factors, Gene Expression Profiling, Hepatocytes, Endothelial Cells, Single-Cell Analysis
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