Engraftment and Repopulation Potential of Late Gestation Fetal Rat Hepatocytes

Boylan, Joan M.; Francois-Vaughan, Heather; Gruppuso, Philip A.; Sanders, Jennifer

doi:10.1097/tp.0000000000001882

Cited by 9 publications

(7 citation statements)

References 39 publications

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“…These data are consistent with studies that demonstrate embryonic day 14 (E14) fetal rat HEPs are still proliferating after 6 months (or equivalent of 12 human years) with full rat liver repopulation (44). Consistent with this, E18 rat F-Heps or phenotypically isolated rat E19 fetal hepatic progenitors were shown more recently to repopulate ~1/3 the liver mass in acute models (45) and in chronic models (46), with evidence of coincident antiinflammatory effects (47). Continued studies are needed to translate fetal liver repopulation to 3D liver organogenesis.…”

Section: Discussionsupporting

confidence: 89%

“…Our data compares favorably to studies of fetal liver proliferative capacity that demonstrate embryonic day 14 (E14) fetal rat hepatocytes (rat F-Heps) are still proliferating after 6 months (or the equivalent of 12 human years) with full rat liver repopulation (Dabeva, Petkov et al 2000). Further, studies of rat E18 F-Heps or phenotypically isolated rat E19 fetal hepatic progenitors were shown more recently to repopulate ∼1/3 the liver mass in acute and chronic models (Boylan, Francois-Vaughan et al 2017);(Bin, Ma et al 2012). Although the mechanisms by which the exponential increase in liver bud growth occurs are not well understood, our study suggests that that fetal hematopoiesis or cellular hypertrophy are not fully responsible.…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Discussionsupporting

confidence: 89%

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

View full text Add to dashboard Cite

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“…Studies demonstrate that hepatic endoderm co-migrates with endothelial cells (Matsumoto, Yoshitomi et al 2001), and STM cells, undergoes branching suggestive of early 3D branching morphogenesis (Watt, Zhao et al 2007), and eventually forms primitive cell sheets consisting of hepatoblasts (Sosa-Pineda, Wigle et al 2000). At later stages of liver organogenesis (E17), recent studies have generally highlighted fetal hepatoblasts proliferation (Boylan, Francois-Vaughan et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Mesenchyme modulates three-dimensional, collective cell migration of liver cellsin vitro- a role for TGFß pathway

Ogoke

Yousef

Ott

et al. 2020

Preprint

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Three dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development/ organogenesis. Here, we modeled liver organogenesis to induce 3D CCM and improve existing models. The liver diverticulum, normally surrounded by septum transversum mesenchyme (STM) at E8.5, was modeled with a miniature liver spheroid surrounded by mesenchymal cells and matrix. In mixed spheroid models with both liver and uniquely MRC5 (fetal lung) fibroblasts, we observed co-migration of cells, and a significant increase in length and number of liver spheroid protrusions, and this was highly sensitive to TGFB1 stimulation. To understand paracrine effects between MRC-5 cells and liver, we performed conditioned medium (M-CM) experiments. Interestingly, the addition of M-CM increased liver 3D CCM, with thin, 3D, dose-dependent branching morphogenesis, an upregulation of Twist1, and a sensitivity to a broad TGFB inhibitor. To test the effects of cell-cell interactions of 3D CCM, the STM was modeled with a spheroid of MRC-5 cells, and we performed co-spheroid culture of liver with MRC-5. We observed a complex morphogenesis, whereby thin, linear, 3D liver cell strands attach to the MRC-5 spheroid, anchor, and thicken to form permanent and thick anchoring contacts between the two spheroids. We also observed spheroid fusion, a form of interstitial migration. In conclusion, we present several novel cultivation systems that induce distinct features of 3D CCM, as judged by the presence of branching, linearity, thickness, and interstitial migration. These methodologies will greatly improve our molecular, cellular, and tissue-scale understanding of liver organogenesis, liver diseases, and liver cell therapy, and will serve as a tool to bridge conventional 2D studies and preclinical in vivo studies.

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“…Ingenuity Pathway Analysis (IPA®), used for pair-wise comparisons, was conducted using only statistically significant probe sets (QIAGEN Inc., https://www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis) [26]. Results with p-values <10 −7 were considered significant based on p-values generated from 5 similarly sized sets of randomly selected genes with a fold-change in expression ≤ 1.1 [27, 28].…”

Section: Methodsmentioning

confidence: 99%

Transcriptional changes during hepatic ischemia-reperfusion in the rat

et al. 2019

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There are few effective targeted strategies to reduce hepatic ischemia-reperfusion (IR) injury, a contributor to poor outcomes in liver transplantation recipients. It has been proposed that IR injury is driven by the generation of reactive oxygen species (ROS). However, recent studies implicate other mediators of the injury response, including mitochondrial metabolic dysfunction. We examined changes in global gene expression after transient hepatic ischemia and at several early reperfusion times to identify potential targets that could be used to protect against IR injury. Male Wistar rats were subjected to 30 minutes of 70% partial warm ischemia followed by 0, 0.5, 2, or 6 hours of reperfusion. RNA was extracted from the reperfused and non-ischemic lobes at each time point for microarray analysis. Identification of differentially expressed genes and pathway analysis were used to characterize IR-induced changes in the hepatic transcriptome. Changes in the reperfused lobes were specific to the various reperfusion times. We made the unexpected observation that many of these changes were also present in tissue from the paired non-ischemic lobes. However, the earliest reperfusion time, 30 minutes, showed a marked increase in the expression of a set of immediate-early genes (c-Fos, c-Jun, Atf3, Egr1) that was exclusive to the reperfused lobe. We interpreted these results as indicating that this early response represented a tissue autonomous response to reperfusion. In contrast, the changes that occurred in both the reperfused and non-ischemic lobes were interpreted as indicating a non-autonomous response resulting from hemodynamic changes and/or circulating factors. These tissue autonomous and non-autonomous responses may serve as targets to ameliorate IR injury.

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Engraftment and Repopulation Potential of Late Gestation Fetal Rat Hepatocytes

Cited by 9 publications

References 39 publications

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

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

Mesenchyme modulates three-dimensional, collective cell migration of liver cellsin vitro- a role for TGFß pathway

Transcriptional changes during hepatic ischemia-reperfusion in the rat

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