Human fetal liver cultures support multiple cell lineages that can engraft immunodeficient mice

Fomin, Marina E.; Beyer, Ashley I.; Muench, Marcus O.

doi:10.1098/rsob.170108

Cited by 27 publications

(16 citation statements)

References 68 publications

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“…Despite strong cytoplasmic autofluorescence, no nuclear 5’ or middle probe staining was identified in the liver; however, 3’ labelling (consistent with dp71) was present ( Extended data , Supplementary figure 5C 60 ). Embryonic liver is host to several distinct cell types (including multiple blood cell lineages 77 ), and interestingly, dp71 labelling appeared restricted to specific nuclei. This isoform might therefore only be expressed in a distinct (minority) population of hepatic cells.…”

Section: Resultsmentioning

confidence: 99%

Single-transcript multiplex in situ hybridisation reveals unique patterns of dystrophin isoform expression in the developing mammalian embryo

Hildyard¹,

Rawson²,

Riddell³

et al. 2020

Wellcome Open Res

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Background: The dystrophin gene has multiple isoforms: full-length dystrophin (dp427) is principally known for its expression in skeletal and cardiac muscle, but is also expressed in the brain, and several internal promoters give rise to shorter, N-terminally truncated isoforms with wider tissue expression patterns (dp260 in the retina, dp140 in the brain and dp71 in many tissues). These isoforms are believed to play unique cellular roles both during embryogenesis and in adulthood, but their shared sequence identity at both mRNA and protein levels makes study of distinct isoforms challenging by conventional methods. Methods: RNAscope is a novel in-situ hybridisation technique that offers single-transcript resolution and the ability to multiplex, with different target sequences assigned to distinct fluorophores. Using probes designed to different regions of the dystrophin transcript (targeting 5', central and 3' sequences of the long dp427 mRNA), we can simultaneously detect and distinguish multiple dystrophin mRNA isoforms at sub-cellular histological levels. We have used these probes in healthy and dystrophic canine embryos to gain unique insights into isoform expression and distribution in the developing mammal. Results: Dp427 is found in developing muscle as expected, apparently enriched at nascent myotendinous junctions. Endothelial and epithelial surfaces express dp71 only. Within the brain and spinal cord, all three isoforms are expressed in spatially distinct regions: dp71 predominates within proliferating germinal layer cells, dp140 within maturing, migrating cells and dp427 appears within more established cell populations. Dystrophin is also found within developing bones and teeth, something previously unreported, and our data suggests orchestrated involvement of multiple isoforms in formation of these tissues. Conclusions: Overall, shorter isoforms appear associated with proliferation and migration, and longer isoforms with terminal lineage commitment: we discuss the distinct structural contributions and transcriptional demands suggested by these findings.

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Section: Resultsmentioning

confidence: 99%

Single-transcript multiplex in situ hybridisation reveals unique patterns of dystrophin isoform expression in the developing mammalian embryo

Hildyard¹,

Rawson²,

Riddell³

et al. 2020

Wellcome Open Res

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“…Foetal liver progenitor cells The foetal liver contains EpCAM+/NCAM+ progenitor cells capable of in vitro expansion and in vivo differentiation into hepatocytes 62,63 (Fig. 2).…”

Section: Alternative Sources Of Hepatocytesmentioning

confidence: 99%

Cell therapy for advanced liver diseases: Repair or rebuild

Dwyer¹,

Macmillan²,

Brennan³

et al. 2021

Journal of Hepatology

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Advanced liver disease presents a significant worldwide health and economic burden and accounts for 3.5% of global mortality. When liver disease progresses to organ failure the only effective treatment is liver transplantation, which necessitates lifelong immunosuppression and carries associated risks. Furthermore, the shortage of suitable donor organs means patients may die waiting for a suitable transplant organ. Cell therapies have made their way from animal studies to a small number of early clinical trials. Herein, we review the current state of cell therapies for liver disease and the mechanisms underpinning their actions (to repair liver tissue or rebuild functional parenchyma). We also discuss cellular therapies that are on the clinical horizon and challenges that must be overcome before routine clinical use is a possibility.

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“…Scientists are particularly interested in modeling liver organogenesis in vivo for disease modeling and therapeutic purposes. Organotypic cultures bearing cell types that are present in the fetal liver, with either primary (42) or stem cell-derived (43) cells, have demonstrated how liver organogenesis can motivate in vivo culture. A key question in liver regenerative medicine is, how does the fetal liver uniquely result in a rapid, massive increase in size?…”

Section: Discussionmentioning

confidence: 99%

High resolution, serial imaging of early mouse and human liver bud morphogenesis in three dimensions

Mon

Ogoke

Shamul

et al. 2019

Preprint

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Liver organogenesis has thus far served as a paradigm for solid organ formation, and has recently attracted interest due to challenges faced in liver regenerative medicine. Murine genetic studies indicate that early steps in morphogenesis are required, suggesting that three-dimensional imaging of early liver morphogenesis at high resolution can improve our understanding. Unfortunately, existing approaches to image early liver morphogenesis have been unable to achieve high spatial resolution (1-5 um) required. In this study, we focused on imaging, visualization, and analysis of early liver development. We utilized available online databases for both mouse (EMAP) and human (3D Atlas of Human Embryology) liver development. To visualize liver bud morphogenesis at high spatial resolution, we performed 3D reconstructions of stacked, digital tissue sections. We show dynamic 3D hepatic cord formation in the mouse in humans. Interestingly, when we quantified fetal liver growth, we showed that 3D fetal liver growth appears to occur in spurts rather continuously, and 3D images suggest that there could be considerable remodeling during these stages. Further, our analysis of the STM, in both mouse and humans, demonstrates that it increases in size during early fetal liver growth, that is highly interconnecting with liver epithelium, and that it can have strong local effects on growth. Finally, we identify and visualize and identify human hepatic cord formation followed by rapid sheet-like growth, which we propose could be an under-appreciated morphological feature that enables rapid growth of early human fetal liver. These studies will motivate future approaches to employ in vitro culture and organoid technology to improve human PSC differentiation, and improve disease modeling, and therapeutic opportunities for liver diseases. In conclusion, compared to 2D sectioning, high spatial resolution imaging of the mouse and human 3D liver bud morphogenesis enables greatly improved visualization of the hepatic cords, 3D sheet-like liver cell growth, STM-epithelial cell interactions, and quantitative comparisons between mouse and human liver bud morphogenesis.

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Human fetal liver cultures support multiple cell lineages that can engraft immunodeficient mice

Cited by 27 publications

References 68 publications

Single-transcript multiplex in situ hybridisation reveals unique patterns of dystrophin isoform expression in the developing mammalian embryo

Single-transcript multiplex in situ hybridisation reveals unique patterns of dystrophin isoform expression in the developing mammalian embryo

Cell therapy for advanced liver diseases: Repair or rebuild

High resolution, serial imaging of early mouse and human liver bud morphogenesis in three dimensions

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