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Human ESMGs show proliferation in association with acinar ductal metaplasia (ADM). (A) A normal ESMG from a human autopsy case without overlying esophageal injury or cancer shows little proliferation at baseline as shown by proliferating cell nuclear antigen staining. (B) An abnormal ESMG from a human esophagectomy case with esophageal adenocarcinoma shows the ductular phenotype within the ESMG with a marked increase in PCNA staining for proliferation.
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Background & Aims
Although cells comprising esophageal submucosal glands (ESMGs) represent a potential progenitor cell niche, new models are needed to understand their capacity to proliferate and differentiate. By histologic appearance, ESMGs have been associated with both overlying normal squamous epithelium and columnar epithelium. Our aim was to...
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Mesenchymal stem cell (MSC)-based liver tissue engineering on nanofibrous scaffold holds great promise for cell-based therapy in liver injuries and end-stage liver failure treatments. MSCs were generated from umbilical cord blood. Hepatogenic differentiation was induced on two-dimensional (2D) and three-dimensional (3D) culture system and character...
Citations
... 45 Furthermore, ESMGs have been isolated and propagated in three-dimensional culture to form 2 types of spheroids, one with squamous markers such as TP63 and one with columnar markers such as KRT7. 46 A truncated form of CAV3 is associated with familial BE and EAC, and although CAV3 is not present in normal squamous esophagus, it is found in rare cells within ESMGs. 82 However, after RFA in porcine esophagus, CAV3 increased in both ESMGs and squamous esophagus, suggesting a role for CAV3 in normal wound healing that may be disrupted in affected patients carrying this truncation. ...
Background & Aims
This review was developed to provide a thorough and effective update on models relevant to esophageal metaplasia, dysplasia, and carcinogenesis, focusing on the advantages and limitations of different models of Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC).
Methods
This expert review was written on the basis of a thorough review of the literature combined with expert interpretation of the state of the field. We emphasized advances over the years 2012–2023 and provided detailed information related to the characterization of established human esophageal cell lines.
Results
New insights have been gained into the pathogenesis of BE and EAC using patient-derived samples and single-cell approaches. Relevant animal models include genetic as well as surgical mouse models and emphasize the development of lesions at the squamocolumnar junction in the mouse stomach. Rat models are generated using surgical approaches that directly connect the small intestine and esophagus. Large animal models have the advantage of including features in human esophagus such as esophageal submucosal glands. Alternatively, cell culture approaches remain important in the field and allow for personalized approaches, and scientific rigor can be ensured by authentication of cell lines.
Conclusions
Research in BE and EAC remains highly relevant given the morbidity and mortality associated with cancers of the tubular esophagus and gastroesophageal junction. Careful selection of models and inclusion of human samples whenever possible will ensure relevance to human health and disease.
... Koltes et al. stated that their model could be used to study inflammation signaling and endotoxin tolerance. Organoids derived from porcine esophageal submucosal glands were derived in 2017 [111]. Two types of organoids were identified in the culture, including hollow/ductal spheroids (characterized by the presence of cytokeratin 7) and solid squamous spheroids (characterized by abundant P63). ...
Preclinical biomedical research is limited by the predictiveness of in vivo and in vitro models. While in vivo models offer the most complex system for experimentation, they are also limited by ethical, financial, and experimental constraints. In vitro models are simplified models that do not offer the same complexity as living animals but do offer financial affordability and more experimental freedom; therefore, they are commonly used. Traditional 2D cell lines cannot fully simulate the complexity of the epithelium of healthy organs and limit scientific progress. The One Health Initiative was established to consolidate human, animal, and environmental health while also tackling complex and multifactorial medical problems. Reverse translational research allows for the sharing of knowledge between clinical research in veterinary and human medicine. Recently, organoid technology has been developed to mimic the original organ’s epithelial microstructure and function more reliably. While human and murine organoids are available, numerous other organoids have been derived from traditional veterinary animals and exotic species in the last decade. With these additional organoid models, species previously excluded from in vitro research are becoming accessible, therefore unlocking potential translational and reverse translational applications of animals with unique adaptations that overcome common problems in veterinary and human medicine.
... And the longitude directions of esophagus are different to the direction of gravity. These differences might affect the severity of esophageal injury caused by BB, but porcine esophagus still represents a superior animal model for human beings (19). ...
Objective
Accidental ingestion of button batteries (BB), usually occurred in children and infants, will rapidly erode the esophagus and result in severe complications, even death. It has been recommended that treatment of this emergent accident as soon as possible with drinking of pH-neutralizing viscous solutions such as honey and sucralfate before surgical removal can mitigate the esophageal injury. Recently, we reported that the electric insulating solutions such as edible oils could mitigate tissue damage in BB-exposed esophageal segments. In this study, we compared the protective effect of kitchen oil with honey or sucralfate, the recommended pH-neutralizing beverages, and with their mixture on esophageal injury caused by BB ingestion in pig esophageal segments and in living piglets.
Methods
Effect of olive oil irrigations was compared to that of honey or sucralfate irrigations in the BB-damaged esophageal segments freshly collected from the local abattoir and in live Bama miniature piglets with the proximal esophagus exposed to BB for 60 min. Also, the effect of olive oil and honey mixture (MOH) irrigations was assessed in live animals. The BB voltage was recorded before insertion and after its removal. Gross and histological analysis of the esophageal injury was performed after BB exposure in segmented fresh esophagus and 7 days after BB exposure in live animals, respectively.
Results
Olive oil irrigations demonstrated better protective effect against BB-induced esophageal damage, compared to honey or sucralfate for BB-induced esophageal damage in vitro . But in vivo study showed that olive oil alone exacerbated esophageal injury because all esophagi irrigated with olive oil perforated. Surprisingly, irrigations with the MOH showed considerable protective effect for BB-induced esophageal damage in live animals, significantly better than irrigations with honey alone. The MOH decreased BB discharge, reduced area of surface injury, attenuated injured depth of esophageal wall thickness, and downed the mucosal injury index in comparison to using honey alone.
Conclusion
Irrigations with olive oil alone couldn’t prevent the BB discharge and is harmful for BB ingestion before surgical removal. However, mixed with honey, olive oil very effectively prevents the BB discharging and produces better esophageal protection than honey.
... In farm animals, porcine intestinal organoid model was first reported using distal duodenum and proximal jejunum of neonates (45). Since then porcine organoid models have been successfully developed from various sections of gastrointestinal tract (49,50) such as esophageal submucosal gland (43), duodenum (23, 51-54), jejunum (23, 46,51,[53][54][55][56][57][58][59][60][61][62][63]66), ileum (23, 44,47,51,53,54,64,65), and the colon (48,51,53,67). Several studies reported bovine organoids derived from jejunum (10, 60), ileum (44,71,73,74), and the colon (72). ...
Animal organoid models derived from farm and companion animals have great potential to contribute to human health as a One Health initiative, which recognize a close inter-relationship among humans, animals and their shared environment and adopt multi-and trans-disciplinary approaches to optimize health outcomes. With recent advances in organoid technology, studies on farm and companion animal organoids have gained more attention in various fields including veterinary medicine, translational medicine and biomedical research. Not only is this because three-dimensional organoids possess unique characteristics from traditional two-dimensional cell cultures including their self-organizing and self-renewing properties and high structural and functional similarities to the originating tissue, but also because relative to conventional genetically modified or artificially induced murine models, companion animal organoids can provide an excellent model for spontaneously occurring diseases which resemble human diseases. These features of companion animal organoids offer a paradigm-shifting approach in biomedical research and improve translatability of in vitro studies to subsequent in vivo studies with spontaneously diseased animals while reducing the use of conventional animal models prior to human clinical trials. Farm animal organoids also could play an important role in investigations of the pathophysiology of zoonotic and reproductive diseases by contributing to public health and improving agricultural production. Here, we discuss a brief history of organoids and the most recent updates on farm and companion animal organoids, followed by discussion on their potential in public health, food security, and comparative medicine as One Health initiatives. We highlight recent evolution in the culturing of organoids and their integration with organ-on-a-chip systems to overcome current limitations in in vitro studies. We envision multidisciplinary work integrating organoid culture and organ-on-a-chip technology can contribute to improving both human and animal health.
... Submucosal glands are tubuloacinar glands that are dispersed throughout the submucosa. They are lined with epithelial cells and mainly secrete mucus, bicarbonate, and growth factors, and their primary function is to lubricate the esophagus and to protect the tissue from the damaging effects of acidic gastric refluxate [4]. Another unique feature of the esophageal wall structure is its mucosa, in which the epithelium has a stratified squamous type with varying thickness and degree of keratinization, which is dependent on the species under investigation. ...
Abstract: Transient receptor potential vanilloid 4 (TRPV4) is a non-selective cation channel that is broadly expressed in different human tissues, including the digestive sys-tem, where it acts as a molecular sensor and a transducer that regulates a variety of functional activities. Despite the extensive research to determine the role of this channel in the physiology and pathophysiology of different organs, the unique morphological and functional features of TRPV4 in the esophagus remain largely unknown. Ten years ago, TRPV4 was shown to be highly expressed in esophageal epithelial cells where its activation induces Ca2+-dependent ATP release, which, in turn, mediates several functions, ranging from mechanosensation to wound healing. This review summarizes the research progress on TRPV4, and focuses on the functional expres-sion of TRPV4 in esophageal epithelium and its possible role in different esophageal diseases that would support TRPV4 as a candidate target for future therapeutic approaches to treat patients with these conditions.
... Although our data support the basal cells in esophageal epithelium as an origin of BE, this study cannot exclude other cells in the esophagus as possible origins of BE. A recent study has shown that the esophageal submucosal gland cells, seen in both human and porcine esophagus, may provide a source of cells to repopulate damaged epithelium in a normal manner (squamous epithelium) or abnormally (columnar epithelium) [48]. In fact, the lack of submucosal glands in rodent esophagus is a limitation of this study and any other studies using rodents. ...
Background
The cellular origin and molecular mechanisms of Barrett’s esophagus (BE) are still controversial. Trans-differentiation is a mechanism characterized by activation of the intestinal differentiation program and inactivation of the squamous differentiation program.AimsRenal capsule grafting (RCG) was used to elucidate whether CDX2 overexpression on the basis of P63 deficiency in the esophageal epithelium may generate intestinal metaplasia.MethodsP63−/−;Villin-Cdx2 embryos were generated by crossing P63+/− mice with Villin-Cdx2 mice. E18.5 esophagus was xenografted in a renal capsule grafting (RCG) model. At 1, 2, or 4 weeks after RCG, the mouse esophagus was immunostained for a proliferation marker (BrdU), squamous transcription factors (SOX2, PAX9), squamous differentiation markers (CK5, CK4, and CK1), intestinal transcription factors (CDX1, HNF1α, HNF4α, GATA4, and GATA6), intestinal columnar epithelial cell markers (A33, CK8), goblet cell marker (MUC2, TFF3), Paneth cell markers (LYZ and SOX9), enteroendocrine cell marker (CHA), and Tuft cell marker (DCAMKL1).ResultsThe P63−/−;Villin-Cdx2 RCG esophagus was lined with proliferating PAS/AB+ cuboidal cells and formed an intestinal crypt-like structure. The goblet cell markers (TFF3 and MUC2) and intestinal transcription factors (CDX1, HNF1α, HNF4α, GATA4, and GATA6) were expressed although no typical morphology of goblet cells was observed. Other intestinal cell markers including enteroendocrine cell marker (CHA), Paneth cell markers (LYZ and Sox9), and intestinal secretory cell marker (UEA/WGA) were also expressed in the P63−/−;Villin-Cdx2 RCG esophagus. Squamous cell markers (PAX9 and SOX2) were also expressed, suggesting a transitional phenotype.ConclusionCDX2 overexpression on the basis of P63 deficiency in esophageal epithelial cells induces Barrett’s-like metaplasia in vivo. Additional factors may be needed to drive this transitional phenotype into full-blown BE.
... Organoids of multiple other organs have also been established. These include canine organoids from the skin (Wiener et al., 2018), prostate gland (Usui et al., 2017), urinary bladder (Elbadawy et al., 2019), kidney and liver (Nantasanti et al., 2015), as well as porcine organoids from the esophagus submucosal gland (von Furstenberg et al., 2017), rectum (Adegbola et al., 2017), and testicle (Sakib et al., 2019;Vermeulen et al., 2019). It also includes those from the mammary gland, pancreatic duct, and endometrium in cattle (Martignani et al., 2018), sheep and horse (Thompson et al., 2020), respectively. ...
Animal models and cell culture in vitro are primarily used in virus and antiviral immune research. Whereas the limitation of these models to recapitulate the viral pathogenesis in humans has been made well aware, it is imperative to introduce more efficient systems to validate emerging viruses in both domestic and wild animals. Organoids ascribe to representative miniatures of organs (i.e., mini-organs), which are derived from three-dimensional culture of stem cells under respective differential conditions mimicking endogenous organogenetic niches. Organoids have broadened virological studies in the human context, particularly in recent uses for COVID19 research. This review examines the status and potential for cross-species applied organotypic culture in validating emerging animal, particularly zoonotic, viruses in domestic and wild animals.
... Figure 7(c) shows the balloon-like structure in the p-H&E image of the lung [48], and the magnified image ( Fig. 7(d)) represents the optically sectioned contour of the alveoli and arteriole clearly. Figure 7(e) shows the squamous epithelium and lamina propria regions of the mucosa of the esophagus tissue distinguished by a basal layer [49], and Fig. 7(f) describes the regions in detail. To obtain high-quality p-H&E images of fresh lung and esophagus specimens, the cutting flatness of the attached side and soft attachment is crucial. ...
A technique using Linnik-based optical coherence microscopy (OCM), with built-in fluorescence microscopy (FM), is demonstrated here to describe cellular-level morphology for fresh porcine and biobank tissue specimens. The proposed method utilizes color-coding to generate digital pseudo-H&E (p-H&E) images. Using the same camera, colocalized FM images are merged with corresponding morphological OCM images using a 24-bit RGB composition process to generate position-matched p-H&E images. From receipt of dissected fresh tissue piece to generation of stitched images, the total processing time is <15 min for a 1-cm² specimen, which is on average two times faster than frozen-section H&E process for fatty or water-rich fresh tissue specimens. This technique was successfully used to scan human and animal fresh tissue pieces, demonstrating its applicability for both biobank and veterinary purposes. We provide an in-depth comparison between p-H&E and human frozen-section H&E images acquired from the same metastatic sentinel lymph node slice (∼10 µm thick), and show the differences, like elastic fibers of a tiny blood vessel and cytoplasm of tumor cells. This optical sectioning technique provides histopathologists with a convenient assessment method that outputs large-field H&E-like images of fresh tissue pieces without requiring any physical embedment.
... This feature is reminiscent of urothelial regenerative processes wherein progenitor cellular hierarchy is modulated by the type of surgical damage encountered 29 . It should also be noted that growth factor dependency in esophageal regenerative processes may also be impacted by physiological differences in the species, sex, and age of the animal model deployed 30,31 . ...
Constructive remodeling of focal esophageal defects with biodegradable acellular grafts relies on the ability of host progenitor cell populations to repopulate implant regions and facilitate growth of de novo functional tissue. Intrinsic molecular mechanisms governing esophageal repair processes following biomaterial-based, surgical reconstruction is largely unknown. In the present study, we utilized mass spectrometry-based quantitative proteomics and in silico pathway evaluations to identify signaling cascades which were significantly activated during neoepithelial formation in a Sprague Dawley rat model of onlay esophagoplasty with acellular silk fibroin scaffolds. Pharmacologic inhibitor and rescue experiments revealed that epithelialization of neotissues is significantly dependent in part on pro-survival stimuli capable of suppressing caspase activity in epithelial progenitors via activation of hepatocyte growth factor receptor (c-MET), tropomyosin receptor kinase A (TrkA), phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt) signaling mechanisms. These data highlight the molecular machinery involved in esophageal epithelial regeneration following surgical repair with acellular implants.
... Later studies applied similar methods with small modifications that were necessary for, or contributed to, the development of enteroids from juvenile and adult pig intestinal tissue [54], or the ability to maintain the enteroids in culture for several months [43,55,56]. Organoids have also been derived successfully from the rectum [57], esophageal submucosal gland [58], and colon (often referred to as "colonoids") [59]. A recent study showed that differential gene-and pathway expression of independent organoid cultures from the same pig was stable over 12 weeks of culture [60]. ...
... In biomedical research on epithelial injury or diseases, porcine organoids may offer possibilities, particularly in situations where collection of tissue samples in humans from the pathogenic site for producing organoids may be too invasive or might induce pathology. Towards this, Von Furstenberg et al. developed an organoid model for porcine esophageal submucosal glands (ESMGs) and showed that the organoid model could be used to study differentiation into squamous versus columnar epithelium, and the mechanisms of ESMG proliferation and regeneration of injured epithelium [58]. Likewise, several studies aimed at providing an animal (porcine) organoid model for (human) biomedical research on small intestinal stem cell physiology and epithelial regeneration in short bowel syndrome, ischemic injury, and other conditions related to intestine [53,54,76]. ...
Organoids are self-organizing, self-renewing three-dimensional cellular structures that resemble organs in structure and function. They can be derived from adult stem cells, embryonic stem cells, or induced pluripotent stem cells. They contain most of the relevant cell types with a topology and cell-to-cell interactions resembling that of the in vivo tissue. The widespread and increasing adoption of organoid-based technologies in human biomedical research is testament to their enormous potential in basic, translational- and applied-research. In a similar fashion there appear to be ample possibilities for research applications of organoids from livestock and companion animals. Furthermore, organoids as in vitro models offer a great possibility to reduce the use of experimental animals. Here, we provide an overview of studies on organoids in livestock and companion animal species, with focus on the methods developed for organoids from a variety of tissues/organs from various animal species and on the applications in veterinary research. Current limitations, and ongoing research to address these limitations, are discussed. Further, we elaborate on a number of fields of research in animal nutrition, host-microbe interactions, animal breeding and genomics, and animal biotechnology, in which organoids may have great potential as an in vitro research tool.
