scholarly journals An Interphase Microfluidic Culture System for the Study of Ex Vivo Intestinal Tissue

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 150 ◽  
Author(s):  
Baydoun ◽  
Treizeibré ◽  
Follet ◽  
Vanneste ◽  
Creusy ◽  
...  
Keyword(s):  
Culture System ◽  
Ex Vivo ◽  
Porous Membrane ◽  
Explant Culture ◽  
Tissue Growth ◽  
Intestinal Tissue ◽  
Microfluidic Perfusion ◽  
Culture Models ◽  
Technical Solutions ◽  
Murine Colon

Ex vivo explant culture models offer unique properties to study complex mechanisms underlying tissue growth, renewal, and disease. A major weakness is the short viability depending on the biopsy origin and preparation protocol. We describe an interphase microfluidic culture system to cultivate full thickness murine colon explants which keeps morphological structures of the tissue up to 192 h. The system was composed of a central well on top of a porous membrane supported by a microchannel structure. The microfluidic perfusion allowed bathing the serosal side while preventing immersion of the villi. After eight days, up to 33% of the samples displayed no histological abnormalities. Numerical simulation of the transport of oxygen and glucose provided technical solutions to improve the functionality of the microdevice.

scholarly journals 3D Organoid Culture From Adult Salivary Gland Tissues as an ex vivo Modeling of Salivary Gland Morphogenesis

2021 ◽  
Vol 9 ◽  
Author(s):  
Donghyun Kim ◽  
Yeo-Jun Yoon ◽  
Dojin Choi ◽  
Jisun Kim ◽  
Jae-Yol Lim
Keyword(s):  
Salivary Gland ◽  
Culture System ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Underlying Mechanism ◽  
Lumen Formation ◽  
Organoid Culture ◽  
Culture Models ◽  
Gland Morphogenesis

Lumen formation of salivary glands has been investigated using in vivo or ex vivo rudiment culture models. In this study, we used a three-dimensional (3D) salivary gland organoid culture system and demonstrated that lumen formation could be recapitulated in mouse SMG organoids. In our organoid culture system, lumen formation was induced by vasoactive intestinal peptide and accelerated by treatment with RA. Furthermore, lumen formation was observed in branching duct-like structure when cultured in combination of fibroblast growth factors (FGF) in the presence of retinoic acid (RA). We suggest RA signaling-mediated regulation of VIPR1 and KRT7 as the underlying mechanism for lumen formation, rather than apoptosis in the organoid culture system. Collectively, our results support a fundamental role for RA in lumen formation and demonstrate the feasibility of 3D organoid culture as a tool for studying salivary gland morphogenesis.

scholarly journals Long-range migration of centrioles to the apical surface of the olfactory epithelium

2021 ◽  
Author(s):  
Kaitlin Ching ◽  
Jennifer T Wang ◽  
Tim Stearns
Keyword(s):  
Olfactory Epithelium ◽  
Culture System ◽  
Sensory Cell ◽  
Ex Vivo ◽  
Explant Culture ◽  
Apical Surface ◽  
Comprehensive Model ◽  
Cell Type ◽  
Range Migration ◽  
Air Liquid Interface

Olfactory sensory neurons (OSNs) in vertebrates detect odorants using multiple cilia, which protrude from the end of the dendrite and require centrioles for their formation. In mouse olfactory epithelium, the centrioles originate in progenitor cells near the basal lamina, often 50 to 100 μm from the apical surface. It is unknown how centrioles traverse this distance or mature to form cilia. Using high-resolution expansion microscopy, we found that centrioles migrate together, with multiple centrioles per group and multiple groups per OSN, during dendrite outgrowth. Centrioles were found by live imaging to migrate slowly, with a maximum rate of 0.18 μm/min. Centrioles in migrating groups were associated with microtubule nucleation factors, but acquired rootletin and appendages only in mature OSNs. The parental centriole had preexisting appendages, formed a single cilium prior to other centrioles, and retained its unique appendage configuration in the mature OSN. We developed an air-liquid interface explant culture system for OSNs and used it to show that centriole migration can be perturbed ex vivo by stabilizing microtubules. We consider these results in the context of a comprehensive model for centriole formation, migration, and maturation in this important sensory cell type.

scholarly journals The Caspase-1/IL-18 Axis of the Inflammasome in Tumor Cells: A Modulator of the Th1/Tc1 Response of Tumor-Infiltrating T Lymphocytes in Colorectal Cancer

Cancers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 189
Author(s):  
Linda Bilonda Mutala ◽  
Cécile Deleine ◽  
Matilde Karakachoff ◽  
Delphine Dansette ◽  
Kathleen Ducoin ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
T Lymphocytes ◽  
Tumor Cells ◽  
Ex Vivo ◽  
Explant Culture ◽  
T Cell Response ◽  
Mrna Levels ◽  
Innate And Adaptive Immunity ◽  
Caspase 1 ◽  
Culture Model

In colorectal cancer (CRC), a high density of T lymphocytes represents a strong prognostic marker in subtypes of CRC. Optimized immunotherapy strategies to boost this T-cell response are still needed. A good candidate is the inflammasome pathway, an emerging player in cancer immunology that bridges innate and adaptive immunity. Its effector protein caspase-1 matures IL-18 that can promote a T-helper/cytotoxic (Th1/Tc1) response. It is still unknown whether tumor cells from CRC possess a functional caspase-1/IL-18 axis that could modulate the Th1/Tc1 response. We used two independent cohorts of CRC patients to assess IL-18 and caspase-1 expression by tumor cells in relation to the density of TILs and the microsatellite status of CRC. Functional and multiparametric approaches at the protein and mRNA levels were performed on an ex vivo CRC explant culture model. We show that, in the majority of CRCs, tumor cells display an activated and functional caspase-1/IL-18 axis that contributes to drive a Th1/Tc1 response elicited by TILs expressing IL-18Rα. Furthermore, unsupervised clustering identified three clusters of CRCs according to the caspase-1/IL-18/TIL density/interferon gamma (IFNγ) axis and microsatellite status. Together, our results strongly suggest that targeting the caspase-1/IL-18 axis can improve the anti-tumor immune response in subgroups of CRC.

ACTIVATED INTESTINAL MUSCLE CELLS PROMOTE PREADIPOCYTE MIGRATION: A NOVEL MECHANISM OF CREEPING FAT FORMATION IN CROHN’S DISEASE

2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S34-S34
Author(s):  
Ren Mao ◽  
Genevieve Doyon ◽  
Ilyssa Gordon ◽  
Jiannan Li ◽  
Sinan Lin ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Ex Vivo ◽  
Muscularis Propria ◽  
Dominant Factor ◽  
Stricture Formation ◽  
Intestinal Tissue ◽  
Mesenteric Fat

Abstract Background and Aims Creeping fat, the wrapping of mesenteric fat around the bowel wall, is a typical feature of Crohn’s disease, and is associated with stricture formation and bowel obstruction. How creeping fat forms is unknown, and we interrogated potential mechanisms using novel intestinal tissue and cell interaction systems. Methods Tissues from normal, ulcerative colitis, non-strictured and strictured Crohn’s disease intestinal specimens were obtained. Fresh and decellularized tissue, mesenteric fat explants, primary human adipocytes, pre-adipocytes, muscularis propria cells, and native extracellular matrix were used in multiple ex vivo and in vitro systems involving cell growth, differentiation and migration, proteomics, and integrin expression. Results Crohn’s disease muscularis propria cells produced an extracellular matrix scaffold which is in direct spatial and functional contact with the immediately overlaid creeping fat. The scaffold contained multiple proteins, but only fibronectin production was singularly upregulated by TGF-b1. The muscle cell-derived matrix triggered migration of pre-adipocytes out of mesenteric fat, fibronectin being the dominant factor responsible for their migration. Blockade of α5β1 on the pre-adipocyte surface inhibited their migration out of mesenteric fat and on 3D decellularized intestinal tissue extracellular matrix. Conclusion Crohn’s disease creeping fat appears to result from the migration of pre-adipocytes out of mesenteric fat and differentiation into adipocytes in response to an increased production of fibronectin by activated muscularis propria cells. These new mechanistic insights may lead to novel approaches for prevention of creeping fat-associated stricture formation.

scholarly journals Scleraxis expressing scleral cells respond to inflammatory stimulation

2021 ◽  
Author(s):  
Ghada Atta ◽  
Falk Schroedl ◽  
Alexandra Kaser-Eichberger ◽  
Gabriel Spitzer ◽  
Andreas Traweger ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Interleukin 1 ◽  
Collagen Fibre ◽  
Tissue Growth ◽  
Collagen Degradation ◽  
Ocular Tissue ◽  
Marker Genes ◽  
Ex Vivo Model ◽  
Tendon Cell ◽  
Fibre Degradation

AbstractThe sclera is an ocular tissue rich of collagenous extracellular matrix, which is built up and maintained by relatively few, still poorly characterized fibroblast-like cells. The aims of this study are to add to the characterization of scleral fibroblasts and to examine the reaction of these fibroblasts to inflammatory stimulation in an ex vivo organotypic model. Scleras of scleraxis-GFP (SCX-GFP) mice were analyzed using immunohistochemistry and qRT-PCR for the expression of the tendon cell associated marker genes scleraxis (SCX), mohawk and tenomodulin. In organotypic tissue culture, explanted scleras of adult scleraxis GFP reporter mice were exposed to 10 ng/ml recombinant interleukin 1-ß (IL1-ß) and IL1-ß in combination with dexamethasone. The tissue was then analyzed by immunofluorescence staining of the inflammation- and fibrosis-associated proteins IL6, COX-2, iNOS, connective tissue growth factor, MMP2, MMP3, and MMP13 as well as for collagen fibre degradation using a Collagen Hybridizing Peptide (CHP) binding assay. The mouse sclera displayed a strong expression of scleraxis promoter-driven GFP, indicating a tendon cell-like phenotype, as well as expression of scleraxis, tenomodulin and mohawk mRNA. Upon IL1-ß stimulation, SCX-GFP+ cells significantly upregulated the expression of all proteins analysed. Moreover, IL1-ß stimulation resulted in significant collagen degradation. Adding the corticosteroid dexamethasone significantly reduced the response to IL1-ß stimulation. Collagen degradation was significantly enhanced in the IL1-ß group. Dexamethasone demonstrated a significant rescue effect. This work provides insights into the characteristics of scleral cells and establishes an ex vivo model of scleral inflammation.

Studies on cellular migration in developing cerebral wall using explant culture system

1989 ◽  
Vol 9 ◽  
pp. 54
Author(s):  
Noriyuki Morita ◽  
Kazuhiro Ikenaka ◽  
Masaharu Ogawa ◽  
Katsuhiko Mikoshiba
Keyword(s):  
Culture System ◽  
Explant Culture ◽  
Cellular Migration

Establishment of a Novel Intervertebral Disc/Endplate Culture Model: Analysis of an Ex Vivo In Vitro Whole-Organ Rabbit Culture System

Spine ◽  
2006 ◽  
Vol 31 (25) ◽  
pp. 2918-2925 ◽  
Author(s):  
Daniel Haschtmann ◽  
Jivko V. Stoyanov ◽  
Ladina Ettinger ◽  
Lutz -P. Nolte ◽  
Stephen J. Ferguson
Keyword(s):  
Intervertebral Disc ◽  
Culture System ◽  
Ex Vivo ◽  
Model Analysis ◽  
Culture Model

scholarly journals An Ex vivo Culture System to Study Thyroid Development

10.3791/51641 ◽  
2014 ◽  
Author(s):  
Anne-Sophie Delmarcelle ◽  
Mylah Villacorte ◽  
Anne-Christine Hick ◽  
Christophe E. Pierreux
Keyword(s):  
Culture System ◽  
Ex Vivo ◽  
Thyroid Development

DESIGN OF AN EX-VIVO VESSEL CULTURE SYSTEM FOR THE STUDY OF EARLY ARTERIALIZATION PHENOMENA IN HUMAN SAPHENOUS BYPASS GRAFTS

2012 ◽  
Vol 45 ◽  
pp. S151
Author(s):  
Marco Piola ◽  
Monica Soncini ◽  
Francesca Prandi ◽  
Maurizio Pesce ◽  
Gianfranco Beniamino Fiore
Keyword(s):  
Culture System ◽  
Ex Vivo ◽  
Bypass Grafts

Establishing an ex vivo culture system for normal pancreatic tissue

Pancreatology ◽  
2014 ◽  
Vol 14 (3) ◽  
pp. S120-S121
Author(s):  
Carlos Fernández Moro ◽  
Sougat Misra ◽  
Soledad Pouso ◽  
Marita Wallenberg ◽  
Rainer Heuchel ◽  
...  
Keyword(s):  
Culture System ◽  
Ex Vivo ◽  
Pancreatic Tissue ◽  
Normal Pancreatic Tissue ◽  
Ex Vivo Culture
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