Personne :
Fradette, Julie

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Structures organisationnelles
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Université Laval. Département de chirurgie
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Voici les éléments 1 - 10 sur 30
  • Publication
    Accès libre
    Oxidative activity of 17β-hydroxysteroid dehydrogenase on testosterone in male abdominal adipose tissues and cellular localization of 17β-HSD type 2
    (North-Holland, 2015-06-26) Fouad Mansour, Mohamed; Boulet, Marie Michèle; Poirier, Donald; Luu The, Van; Brochu, Gaétan; Cianflone, Katherine M.; Lebel, Stefane; Pelletier, Mélissa; Fradette, Julie; Tchernof, André; Mayrand, Dominique
    Testosterone can be converted into androstenedione (4-dione) by 17β-hydroxysteroid dehydrogenase (HSD) activity likely performed by 17β-HSD type 2. Our objective was to evaluate the rate of testosterone conversion to 4-dione as well as expression and localization of 17β-HSD type 2 in omental (OM) vs. subcutaneous (SC) adipose tissues of men. Formation of 4-dione from testosterone was significantly higher in homogenates (p ≤ 0.001) and explants (p ≤ 0.01) of OM than SC tissue. Microscopy analyses and biochemical assays in cell fractions localized the enzyme in the vasculature/endothelial cells of adipose tissues. Conversion of testosterone to 4-dione was weakly detected in most OM and/or SC preadipocyte cultures. Positive correlations were found between 17β-HSD type 2 activity in whole tissue and BMI or SC adipocyte diameter. We conclude that conversion of testosterone to 4-dione detected in abdominal adipose tissue is caused by 17β-HSD type 2 which is localized in the vasculature of the adipose compartment.
  • Publication
    Accès libre
    Creating capillary networks within human engineered tissues: impact of adipocytes and their secretory products
    (Elsevier, 2014-09-30) Vincent, Caroline; Proulx, Maryse; Aubin, Kim; Fradette, Julie; Mayrand, Dominique
    The development of tissue-engineered substitutes of substantial volume is closely associated with the need to ensure rapid vascularization upon grafting. Strategies promoting angiogenesis include the in vitro formation of capillary-like networks within engineered substitutes. We generated both connective and adipose tissues based on a cell sheet technology using human adipose-derived stromal cells. This study evaluates the morphology and extent of the capillary networks that developed upon seeding of human microvascular endothelial cells during tissue production. We posited that adipocyte presence/secretory products could modulate the resulting capillary network when compared to connective substitutes. Analyses including confocal imaging of CD31-labeled capillary-like networks indicated slight differences in their morphological appearance. However, the total volume occupied by the networks as well as the frequency distribution of the structure’s volumes were similar between connective and adipose tissues. The average diameter of the capillary structures tended to be 20% higher in reconstructed adipose tissues. Quantification of pro-angiogenic molecules in conditioned media showed greater amounts of leptin (15×), angiopoietin-1 (3.4×) and HGF (1.7×) secreted from adipose than connective tissues at the time of endothelial cell seeding. However, this difference was attenuated during the following coculture period in endothelial cell-containing media, correlating with the minor differences noted between the networks. Taken together, we developed a protocol allowing reconstruction of both connective and adipose tissues featuring well-developed capillary networks in vitro. We performed a detailed characterization of the network architecture within engineered tissues that is relevant for graft assessment before implantation as well as for in vitro screening of angiogenic modulators using three-dimensional models.
  • Publication
    Stem cells of the skin and cornea : their clinical applications in regenerative medicine
    (Rapid Science Publishers, 2011-02-01) Germain, Lucie; Larouche, Danielle; Gauvin, Robert; Proulx, Stéphanie; Fradette, Julie
    Purpose of review: The use of stem cells is of great interest for the treatment of various pathologies and ultimately for the restoration of organ function. Progress pointing towards future treatments of skin and corneal epithelial stem cell defects are reviewed, including the transplantation of living tissue-engineered substitutes. Recent findings: This article focuses on substitutes optimized for permanent replacement of skin and cornea. New skin substitutes for burn care are currently under development. More complex tissue-engineered skin substitutes in which stroma, adipose tissue, capillaries, and neurons are combined with the epithelium are being developed. Some dermal/epidermal substitutes have been applied to the treatment of patients. Cultured corneal epithelial cells have been characterized and more complete corneal substitutes are being designed. Long-term clinical results on the transplantation of cultured corneal stem cells for the treatment of limbal stem cell deficiency have been reported. Summary: Advances in tissue engineering for the development of substitutes that will benefit patients suffering from skin or corneal stem cell deficiencies are reviewed. These products are often a combination of cells, scaffolds and other factors. Key considerations in the development of corneal and skin substitutes for clinical applications are discussed.
  • Publication
    Localization of merkel cells at hairless and hairy human skin sites using keratin 18
    (National Research Council of Canada, 1995-09-01) Michel, Martine; Germain, Lucie; Godbout, Marie-Josée; Fradette, Julie
    Les cellules de Merkel sont des cellules cutanées particulières, possédant non seulement des granules de sécrétion contenant des neuropeptides, mais également des desmosomes et des tonofilaments formés des kératines 8, 18, 19 et 20. Les cellules de Merkel sont plutôt rares dans la peau d'adulte. Toutefois, elles ont été observées en plus grande quantité dans les régions sans poils, telles la paume des mains et la plante des pieds. Leur présence a aussi été rapportée dans les follicules pileux. Les cellules de Merkel sont souvent innervées par des fibres nerveuses sensorielles et on leur reconnaît un rôle de mécanorécepteurs dans la peau. Toutefois, leur origine et fonction précises ne sont pas encore clairement établies. Le but de cette étude était de localiser les cellules de Merkel dans des régions anatomiques pourvues ou non de follicules pileux, par immunohistochimie avec les anticorps Ks18.174 et Ks19.1 respectivement dirigés contre les kératines 18 et 19. Dans la paume des mains et la plante des pieds, les cellules de Merkel ont été identifiées dans la couche basale de l'épiderme, à la base des papilles. Pour étudier la localisation des cellules de Merkel dans une région possédant des poils, nous avons choisi la peau provenant de réduction mammaire car on y trouve de petits follicules pileux représentatifs des poils couvrant une grande partie de notre corps. Des cellules de Merkel ont été observées dans l'épiderme interfolliculaire et dans le follicule pileux, où elles ont été localisées dans la région de l'isthme.Mots clés : peau, humain, cellule de Merkel, kératines, follicule pileux.
  • Publication
    Vibrissa hair bulge houses two populations of skin epithelial stem cells distinct by their keratin profile
    (Federation of American Societies for Experimental Biology, 2007-12-27) Germain, Lucie; Tong, Xuemei; Larouche, Danielle; Fradette, Julie; Coulombe, Pierre A.
    Defining the properties of postnatal stem cells is of interest given their relevance for tissue homeostasis and therapeutic applications, such as skin tissue engineering for burn patients. In hair follicles, the bulge region of the outer root sheath houses stem cells. We show that explants from the prominent bulge area, but not the bulb, in rodent vibrissa follicles can produce epidermis in a skin model of tissue engineering. Using morphological criteria and keratin expression, we typified epithelial stem cells of vibrissa bulge. Two types of slow-cycling cells (Bb, Bs1) featuring a high colony-forming capacity occur in the bulge. Bb cells are located in the outermost basal layer, express K5, K15, K17, and K19, and feature a loosely organized keratin network. Bs1 cells localize to the suprabasal layers proximal to Bb cells and express K5/K17, corre lating with a network of densely bundled filaments. These prominent bundles are missing in K17-null mice, which lack vibrissa. Atypically, both the Bb and Bs1 keratinocytes lack K14 expression. These findings show heterogeneity within the hair follicle stem cell reposi tory, establish that a subset of slow-cycling cells are suprabasal in location, and point to a special role for K5/K17 filaments in a newly defined subset of stem cells. Our results are discussed in the context of long-term survival of engineered tissues after grafting that requires the presence of stem cells.
  • Publication
    Accès libre
    Characterization of in vitro engineered human adipose tissues : relevant adipokine secretion and impact of TNF-α
    (Public Library of Science, 2015-09-14) Roy, Alphonse; Proulx, Maryse; Côté, Jean-François; Safoine, Meryem; Aubin, Kim; Audet-Casgrain, Marie-Alice; Fradette, Julie; Têtu, Félix-André
    Representative modelling of human adipose tissue functions is central to metabolic research. Tridimensional models able to recreate human adipogenesis in a physiological tissue-like context in vitro are still scarce. We describe the engineering of white adipose tissues reconstructed from their cultured adipose-derived stromal precursor cells. We hypothesize that these reconstructed tissues can recapitulate key functions of AT under basal and pro-inflammatory conditions. These tissues, featuring human adipocytes surrounded by stroma, were stable and metabolically active in long-term cultures (at least 11 weeks). Secretion of major adipokines and growth factors by the reconstructed tissues was determined and compared to media conditioned by human native fat explants. Interestingly, the secretory profiles of the reconstructed adipose tissues indicated an abundant production of leptin, PAI-1 and angiopoietin-1 proteins, while higher HGF levels were detected for the human fat explants. We next demonstrated the responsiveness of the tissues to the pro-inflammatory stimulus TNF-α, as reflected by modulation of MCP-1, NGF and HGF secretion, while VEGF and leptin protein expression did not vary. TNF-α exposure induced changes in gene expression for adipocyte metabolism-associated mRNAs such as SLC2A4, FASN and LIPE, as well as for genes implicated in NF-κB activation. Finally, this model was customized to feature adipocytes representative of progressive stages of differentiation, thereby allowing investigations using newly differentiated or more mature adipocytes. In conclusion, we produced tridimensional tissues engineered in vitro that are able to recapitulate key characteristics of subcutaneous white adipose tissue. These tissues are produced from human cells and their neo-synthesized matrix elements without exogenous or synthetic biomaterials. Therefore, they represent unique tools to investigate the effects of pharmacologically active products on human stromal cells, extracellular matrix and differentiated adipocytes, in addition to compounds modulating adipogenesis from precursor cells.
  • Publication
    High definition confocal imaging modalities for the characterization of tissue-engineered substitutes
    (Springer, 2018-04-01) Fradette, Julie; Mayrand, Dominique
    Optimal imaging methods are necessary in order to perform a detailed characterization of thick tissue samples from either native or engineered tissues. Tissue-engineered substitutes are featuring increasing complexity including multiple cell types and capillary-like networks. Therefore, technical approaches allowing the visualization of the inner structural organization and cellular composition of tissues are needed. This chapter describes an optical clearing technique which facilitates the detailed characterization of whole-mount samples from skin and adipose tissues (ex vivo tissues and in vitro tissue-engineered substitutes) when combined with spectral confocal microscopy and quantitative analysis on image renderings.
  • Publication
    Considerations in the choice of a skin donor site for harvesting keratinocytes containing a high proportion of stem cells for culture in vitro
    (Butterworth-Heinemann, 2010-12-03) Germain, Lucie; Larouche, Danielle; Paquet, Claudie; Fugère, Claudia.; Genest, Hervé; Auger, François A.; Gauvin, Robert; Têtu, Félix-Andre; Bouchard, Maurice; Roy, Aphonse; Fradette, Julie; Lavoie, Amélie; Beauparlant, Annie.
    The treatment of severely burned patients has benefited from the grafting of skin substitutes obtained by expansion of epithelial cells in culture. The aim of this study was to evaluate whether the anatomic site chosen for harvesting skin had an impact on the quality of the derived cell cultures. Considering that hair follicles contain epithelial stem cells, we compared hairy skin sites featuring different densities and sizes of hair follicles for their capacity to generate high quality keratinocyte cultures. Three anatomic sites from adult subjects were compared: scalp, chest skin and p-auricular (comprising pre-auricular and post-auricular) skin. Keratin (K) 19 was used as a marker to evaluate the proportion of stem cells. Keratinocytes were isolated using the two-step thermolysin and trypsin cell extraction method, and cultured in vitro. The proportion of K19-positive cells harvested from p-auricular skin was about twice that of the scalp. This K19-positive cell content also remained higher during the first subcultures. In contrast to these in vitro results, the number of K19-positive cells estimated in situ on skin sections was about double in scalp as in p-auricular skin. Chest skin had the lowest number of K19-positive cells. These results indicate that in addition to the choice of an adult anatomic site featuring a high number of stem cells in situ, the quality of the cultures greatly depends on the ability to extract stem cells from the skin biopsy
  • Publication
    Accès libre
    Magnetic resonance imaging of human tissue-engineered adipose substitutes
    (Mary Ann Liebert, Inc, 2015-02-23) Audet, Pierre; Proulx, Maryse; Auger, Michèle; Fortin, Marc-André; Aubin, Kim; Lagueux, Jean; Fradette, Julie
    Adipose tissue (AT) substitutes are being developed to answer the strong demand in reconstructive surgery. To facilitate the validation of their functional performance in vivo, and to avoid resorting to excessive number of animals, it is crucial at this stage to develop biomedical imaging methodologies, enabling the follow-up of reconstructed AT substitutes. Until now, biomedical imaging of AT substitutes has scarcely been reported in the literature. Therefore, the optimal parameters enabling good resolution, appropriate contrast, and graft delineation, as well as blood perfusion validation, must be studied and reported. In this study, human adipose substitutes produced from adipose-derived stem/stromal cells using the self-assembly approach of tissue engineering were implanted into athymic mice. The fate of the reconstructed AT substitutes implanted in vivo was successfully followed by magnetic resonance imaging (MRI), which is the imaging modality of choice for visualizing soft ATs. T1-weighted images allowed clear delineation of the grafts, followed by volume integration. The magnetic resonance (MR) signal of reconstructed AT was studied in vitro by proton nuclear magnetic resonance (1H-NMR). This confirmed the presence of a strong triglyceride peak of short longitudinal proton relaxation time (T1) values (200±53 ms) in reconstructed AT substitutes (total T1=813±76 ms), which establishes a clear signal difference between adjacent muscle, connective tissue, and native fat (total T1 ∼300 ms). Graft volume retention was followed up to 6 weeks after implantation, revealing a gradual resorption rate averaging at 44% of initial substitute's volume. In addition, vascular perfusion measured by dynamic contrast-enhanced-MRI confirmed the graft's vascularization postimplantation (14 and 21 days after grafting). Histological analysis of the grafted tissues revealed the persistence of numerous adipocytes without evidence of cysts or tissue necrosis. This study describes the in vivo grafting of human adipose substitutes devoid of exogenous matrix components, and for the first time, the optimal parameters necessary to achieve efficient MRI visualization of grafted tissue-engineered adipose substitutes.
  • Publication
    Human adipose-derived stromal cells for the production of completely autologous self-assembled tissue-engineered vascular substitutes
    (Minerals, Metals and Materials Society, 2015-06-15) Tondreau, Maxime; Laterreur, Véronique; Germain, Lucie; Vallières, Karine; Ruel, Jean; Auger, François A.; Fradette, Julie
    There is a clinical need for small-diameter vascular substitutes, notably for coronary and peripheral artery bypass procedures since these surgeries are limited by the availability of grafting material. This study reports the characterization of a novel autologous tissue-engineered vascular substitute (TEVS) produced in 10weeks exclusively from human adipose-derived stromal cells (ASC) self-assembly, and its comparison to an established model made from dermal fibroblasts (DF). Briefly, ASC and DF were cultured with ascorbate to form cell sheets subsequently rolled around a mandrel. These TEVS were further cultured as a maturation period before undergoing mechanical testing, histological analyses and endothelialization. No significant differences were measured in burst pressure, suture strength, failure load, elastic modulus and failure strain according to the cell type used to produce the TEVS. Indeed, ASC- and DF-TEVS both displayed burst pressures well above maximal physiological blood pressure. However, ASC-TEVS were 1.40-fold more compliant than DF-TEVS. The structural matrix, comprising collagens type I and III, fibronectin and elastin, was very similar in all TEVS although histological analysis showed a wavier and less dense collagen matrix in ASC-TEVS. This difference in collagen organization could explain their higher compliance. Finally, human umbilical vein endothelial cells (HUVEC) successfully formed a confluent endothelium on ASC and DF cell sheets, as well as inside ASC-TEVS. Our results demonstrated that ASC are an alternative cell source for the production of TEVS displaying good mechanical properties and appropriate endothelialization.