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Academic literature on the topic 'Cellules stromales mésenchymateuses de la gelée de Wharton'
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Dissertations / Theses on the topic "Cellules stromales mésenchymateuses de la gelée de Wharton"
Reppel, Loïc. "Potentialité des cellules stromales de la gelée de Wharton en ingénierie du cartillage." Thesis, Université de Lorraine, 2014. http://www.theses.fr/2014LORR0164/document.
Full textMesenchymal Stromal/Stem Cells from human Wharton’s jelly (WJ-MSC) are an abundant and interesting source of stem cells for applications in cell and tissue engineering. Their fetal origin confers specific characteristics compared to Mesenchymal Stromal/Stem Cells isolated from human bone marrow (BM-MSC). First, the aim of this work is to optimize WJ-MSC culture conditions for their subsequent clinical use. We focus on the influence of oxygen concentration during monolayer expansion on several parameters to characterize MSC. The results are compared to those obtained with BM-MSC. Our work distinguishes WJ-MSC from BM-MSC in terms of proliferation and adipogenic differentiation. Considering our results, hypoxia during cell expansion is an important parameter to take into account regarding proliferation potential but also chondrogenic differentiation potential. The influence of obstetric factors on WJ-MSC characteristics is also explored. In cartilage tissue engineering context, the second phase of the project is to induce cell differentiation into chondrocytes by seeding them in Alginate/Hyaluronic Acid hydrogel scaffold, and during 28 days. The results obtained are compared to those obtained with BM-MSC. After 4 weeks of culture, WJ-MSC are able to adapt to their environment and express specific cartilage-Related genes and matrix proteins such as type 2 collagen, which is found more expressed after differentiation fromWJ-MSC, than from BM-MSC
Margossian, Talar. "Caractérisation des cellules souches mésenchymateuses du sang placentaire et de la gelée de Wharton." Thesis, Université de Lorraine, 2013. http://www.theses.fr/2013LORR0030/document.
Full textStem cells are the hopes for cell therapy and tissue engineering. MSCs from fetal tissue (umbilical cord blood and WJ), which are a source of embryonic epiblast grow relatively faster comparing to other adult sources. The culture condition can affect cell behavior. In our study, we explored the effect of oxygen concentration on the expansion, immunophenotyping, and differentiation of these cells. The aim of this work is to identify the optimal method for isolation of MSCs derived from fetal tissue. Given the low rate of success in the isolation of MSCs from cord blood, we headed to WJ-MSCs. We have determined in siu, the specific markers expressed in the WJ and in the perivascular region. Studies on the morphology growth kinetics, and phenotypic expression of surface makers of MSCs isolated from WJ were made over a long period (7 passages) in different culture conditions. We have shown that WJ is composed of an abundant extracellular matrix rich in collagen and glycasominoglycans and have variable phenotype depending from their localization in the jelly. This tissue is able to provide a large amount of MSCs (6.7x105 Cs/cm of cord) that maintain a constant morphology. Finally, regardless of the passage, the oxygen concentration does not effect on the phenotype of the cells. In contrast, a low oxygen concentration during expansion appears to decrease the doubling time of MSCs, promote chondrogenesis and inhibit osteogenic differentiation. Finally, whatever the culture conditions, adipogenic differentiation of WJ-MSC seems difficult to obtain
Ye, Junsong. "Ingénierie tissulaire hépatique à partir du foie décellularisé et de cellules souches mésenchymateuses de la gelée de Wharton." Thesis, Université de Lorraine, 2015. http://www.theses.fr/2015LORR0127/document.
Full textThere are over 100 forms of liver diseases caused by various factors and affecting a lot of people. Unfortunately, the only treatment of a terminal liver disease is liver transplantation. However, liver transplantation often fails because of the deficit in human liver donors. Recently, a new innovative alternative for treating end-stage liver disease appears: self-built organ. In liver tissue engineering the source of cells, the decellularized liver scaffold and circular culture bioreactor, are essential factors to be taken into account. The objective of this thesis is to study two steps needed for the development of an artificial liver : cells and organ decellularization. In the first stage, we collected and characterize Wharton’s-Jelly mesenchymal stem cells (WJ-MSCs), and their differentiation potential into hepatocytes. In the second stage of the work, we developed a method for liver decellularization. We were able to get acellular scaffolds by continuous perfusion with 1% SDS and Triton X100 1%. In conclusion, this study shows the capability of WJ-MSC to be differentiated into hepatocytes and the feasibility to obtain acellular livers. That open perspectives toward the development of an artificial liver and the treatment of liver diseases
Avercenc-Léger, Léonore. "Devenir des propriétés immunomodulatrices des cellules souches mésenchymateuses de la gelée de Wharton au cours de la différenciation chondrocytaire." Thesis, Université de Lorraine, 2017. http://www.theses.fr/2017LORR0210/document.
Full textThe purpose of this work is to determine the optimal conditions for allogeneic substitutes production, adapted to filling the cartilaginous lesions in osteoarthritis treatment. It focuses on the cellular component of these substitutes. The use of mesenchymal stem cells from umbilical cords (WJ-MSC) involves determining which factors, related to direct and indirect environment of the WJ-MSC, can influence their proliferation and chondrogenic differentiation. In a first part of our work, three types of factors were studied: related to the donor child, the course of labor and delivery, pregnancy and the mother. Our results show that WJ-MSC have enhanced proliferative capacities when coming from full-term birth and without complications, with the use of Syntocinon® during labor. On this basis, we used the most effective WJ-MSC for cartilage engineering. It was then essential to elucidate their action profile in allogeneic context. We stimulated WJ-MSC embedded in Alginate/Hyaluronic Acid (Alg/HA) scaffolds with different concentrations of IFN-γ and TNF-α in order to determine the most effective stimulation profile, with regard to viability of the cells and evolution of immunomodulatory soluble factors secretion. According to our results, the stimulation by IFN-γ and TNF-α on WJ-MSC in Alg/HA scaffolds is more effective when these two cytokines are used together and is not deleterious for cell viability at the concentrations of 20 and 30 ng/mL, respectively. This double stimulation induces an increase in the secretion of IL-6 and PGE-2 by the WJ-MSC, a decrease in the secretion of VEGF and does not modify the secretion of TGF-β. We confirmed these data during a functional study: cocultures with peripheral blood mononuclear cells (PBMC) from healthy donors allowed us to evaluate the response of WJ-MSC in an allogeneic situation. These allogeneic situations have been studied at different times to evaluate the immunological properties of WJ-MSC during the time of chondrogenic differentiation. Our results show that WJ-MSC can express HLA-G molecules as well as IDO, but these expressions are limited in Alg/HA biomaterials. Finally, the WJ-MSC in Alg/HA biomaterials in allogeneic conditions are not immunogenic, regardless of the time of differentiation. On the other hand, their immunomodulatory capacities decrease over time and are stronger at day 0 and day 3 of chondrogenic differentiation, which leads to an early use of these cells. Finally, this work allows us to (i) select the umbilical cords suitable for cellular and cartilage engineering, (ii) define the conditions mimicking in vitro an allogeneic situation, (iii) elucidate the immunomodulatory properties of WJ-MSC during Alg/HA biomaterials chondrogenic differentiation, including allogeneic situations
Omar, Reine El. "Étude de l'effet de la différention endothéliale sur les propriétés immunomodulatrices des cellules souches mésenchymateuses issues de la gelée de Wharton du cordon ombilical humain." Thesis, Université de Lorraine, 2014. http://www.theses.fr/2014LORR0126/document.
Full textUmbilical cord Wharton’s jelly mesenchymal stem cells (WJ-MSC) are immune-privileged and immunosuppressive. Our study sought to determine the effect of endothelial differentiation on the immunomodulatory capacities of WJ-MSC. Endothelial-like cells (ECL) differentiation was performed by seeding MSC on polyelectrolyte multilayer films as substrate and stimulating them by EGM-2 culture medium. The expression of two immunological markers HLA-DR and CD86 was followed during the differentiation time. The effect of co-culture with ELC or MSC either in contact or separated by a transwell on different immune cells (peripheral blood mononuclear cells, T Lymphocytes (LT), Natural Killer (NK) cells) was assessed by evaluating immune cells proliferation. The results showed that ELC expressed endothelial markers, expressed low level of HLA-DR and CD86 and inhibited the proliferation of the different immune cell populations, and this inhibition is thought to be mediated by soluble factors as IDO, IL-6, IL-1β and PGE2, and by direct cellular contact. We reported also the capacity of ELC to generate a population of regulatory T cells and to decrease the expression of activating receptor NKG2D by NK cells. Moreover, we demonstrated that co-cultures with ELC induce CD73 expression on NK cells, a mechanism that may induce adenosine (a potent immunosuppressor) secretion by NK cells. Thus, ELC maintain the hypo-immunogenic and immunosuppressive characters of WJ-MSC suggesting their possible use in vascular engineering
Laroye, Caroline. "Le cordon ombilical : une source alternative de cellules souches/stromales mésenchymateuses dans le traitement du choc septique ?" Thesis, Université de Lorraine, 2017. http://www.theses.fr/2017LORR0279.
Full textSeptic shock, equal to the myocardial infraction, is currently the tenth cause of death in the world. The pathophysiological complexity of this syndrome, with a simultaneous pro and anti- inflammatory state, results in the failure of conventional treatments. In this sense, research is focusing on innovative therapeutics agent, including mesenchymal stem cells (MSC). Indeed, murine studies of septic shock showed that MSC improve organ injuries, bacteremia and survival by notably a paracrine mechanism. However, MSC properties vary according to the source tissue, especially if they are derived from a fetal tissue (Wharton’s jelly (WJ), placenta amniotic fluid) or an adult tissue (bone marrow (BM), adipose tissue...). Our first objective was to compare, in a septic shock murine model, the effect of BM-MSC with that of WJ-MSC. Although some differences were observed, the same efficiency was demonstrated between these two sources. However, WJ-MSC present large advantages in comparison to BM-MSC due to their important proliferation capacities and potential quantities of umbilical cord donation. Consequently, our second objective was to investigate the effect of WJ-MSC administration in a relevant pig model of peritonitis in order to better mimic a clinical approach in humans. This study, conducted in double-blind and in presence of an experimented intensivist, showed that WJ-MSC produced in clinical grade and used immediately after thawing, improve survival, hemodynamic parameters and organ injuries by another action than that described in murine studies
Degoul, Olivier. "Optimisation des techniques de traitement des échantillons de cordon ombilical et immunophénotypage multiparamétrique des cellules souches mésenchymateuses de la gelée de Wharton du cordon ombilical pour des applications en thérapie cellulaire." Thesis, Lyon 1, 2014. http://www.theses.fr/2014LYO10174.
Full textWith around 136,000,000 births per year worldwide stem cells isolated from neonatal tissues such as the umbilical cord have many advantages in term of access and availability. The umbilical cord contains a connective tissue present in large quantities surrounding the blood vessels, the Wharton’s jelly, demonstrated as a very rich source of Mesenchymal Stem Cells (MSCs). Those multipotent stem cells isolated from the umbilical cord have shown self‐renewal and differentiation capability into a range of cells inherent potential for cell therapy applications not only as differentiated cells but also thanks to potent capabilities for immunomodulation and regulation of inflammation. In this work we designed process and tools allowing efficient use of umbilical cord samples and the isolation of standardized Wharton's Jelly tissue explants. We studied techniques improving MSC isolation from tissue explants and developed an in‐vitro culture protocol, free of Foetal Bovine Serum (FBS) and other animal sourced products and thus more likely to be used in clinical settings. We developed a multiparametric flow cytometry protocol dedicated to phenotyping the surface expression of 27 different antigens present on Wharton's Jelly MSCs and to study the effect of several in‐vitro culture conditions and events on their surface phenotype. The multiparametric protocol was used to compare surface expression of MSCs isolated from Wharton's jelly, bone marrow and skin harvested fibroblasts. Finally, clinical development of human WJ‐MSC in a bone tissue differentiation protocol was evaluated for cleft palate repair, demonstrating the useful application of this tissue source
Lesieur, Romane. "Ingénierie tissulaire de l'oesophage." Electronic Thesis or Diss., Bordeaux, 2024. http://www.theses.fr/2024BORD0020.
Full textUpon removal of a portion of the esophagus, the restoration of the digestive continuity involves the surgical creation of an intrathoracic esophagogastric anastomosis. However, postoperative complications such as lung impairments, fistulas, strictures, graft necrosis, and gastroesophageal reflux are reported. The enhancement of surgical procedures for esophageal replacement has made promising progress by the development of a substitute through tissue engineering that utilizes a decellularized biological esophageal matrix (DEM). The primary objective of this study was to optimize the design of porcine DEM and characterize its biological and mechanical properties. The secondary objective was to cellularize DEM using readily available immune-privileged human mesenchymal stromal cells derived from Wharton's jelly (hMSCs-WJ).Esophageal decellularization was performed according to a protocol based on the dynamic perfusion of chemical and enzymatic solutions through the organ lumen. Histological analysis and residual DNA quantification of the DEM were conducted to determine the efficiency of the decellularization protocol. The ultrastructure of the DEM was analyzed using immunohistochemical (IHC) labeling, and the composition of the extracellular matrix (ECM) protein content was described by mass spectrometry. In-vitro cytotoxicity tests of DEM were conducted following ISO 10993-5 standards. The evaluation of suture retention strength, tensile strength, and bursting pressure of DEM aimed to describe the mechanical behavior of the substitute for clinical use.hMSCs-WJ used for DEM cellularization were extracted from human umbilical cords, and their flow cytometry profiling confirmed the purity of the cell population. The immune response of hMSCs-WJ was quantified after co-culture with peripheral blood mononuclear cells (PBMCs). PBMCs phenotyping assessed the expression of immune markers in contact with hMSCs-WJ, while enzyme-linked immunosorbent assay (ELISA) quantified cytokine release. The proposed DEM cellularization strategy involved the development of cell sheets from hMSCs-WJ. The validation of the cell sheet production protocol involved the characterization of the cellular phenotype by IHC analysis, and the mechanical study of the sheets measured their resistance to perforation.The absence of cellular content and residual DNA quantification in DEM confirmed the efficacy of decellularization according to current validation criteria. The ultrastructure and biological components of the ECM were preserved, and proteomic analysis highlighted protein complexity. Decellularization treatment did not induce DEM toxicity, and the mechanical behavior of DEM was suitable for its use as an esophageal substitute.Culturing hMSCs-WJ as cell sheets promoted the cellularization of the DEM. Once seeded, the sheets retained their cellular phenotype and immune-privileged characteristics. In-vitro tissue remodeling was visible, along with the formation of a new ECM produced by hMSCs-WJ.Characterization of the obtained DEM offered biological complexity and favorable mechanical behavior for its use as an esophageal substitute. DEM was cellularizable with hMSCs-WJ cell sheets, potentially promoting tissue integration and remodeling
Rammal, Hassan. "Comparaison de la capacité de différenciation en cellules endothéliales, de deux types de cellules souches mésenchymateuses issues de la gelée de Wharton et de la moelle osseuse." Thesis, Université de Lorraine, 2014. http://www.theses.fr/2014LORR0008/document.
Full textThe incidence of cardiovascular disease remains a major public health problem. Despite the development of endovascular therapies, surgical treatment is necessary for many patients. The low availability of natural vessels, autologous or not, and the mechanical and biological limits of artificial substitutes, led to the use of a new domain: vascular engineering. In recent years, the concept has emerged and evolved. He could afford to offer new types of synthetic vascular substitutes and / or biological, in particular through the use of stem cells, offering interesting perspectives in the field of vascular engineering. The purpose of this study was to obtain a reliable and reproducible protocol to generate functional endothelial cells (ECs) from mesenchymal stem cells (MSCs) derived from the umbilical cord Wharton's jelly (WJ) and bone marrow (BM). Nevertheless, their differentiation into vascular cells needs a culture surface functionalization; our group demonstrated the potential use of polyelectrolyte multilayer made of poly(allylamine hydrochloride): PAH, and poly(styrene sulfonate): PSS, in promoting cells adhesion, proliferation and differentiation. Cells were cultured on (PAH-PSS)4 films or collagen type I (used as control), in the presence of angiogenic growth factors. Cells differentiation into EC was followed through the expression of endothelial markers (PCR and western blot); cell functionality was checked through their ability to incorporate acetylated LDL (Low Density Lipoprotein), to produce NO (Nitric oxide) and to express the von Willebrand factor (vWF). After 14 days of stimulation, only WJ-MSCs were able to generate functional ECs demonstrating the potential of combining WJ-MSCs and (PAH-PSS)4 films in vascular tissue engineering field
Harmouch, Chaza. "Évaluation de l'adhésion et de la différenciation endothéliale de cellules souches mésenchymateuses issues de la gelée de Wharton du cordon ombilical humain sur des supports fonctionnalisés du type chitosane/hyaluronane." Thesis, Université de Lorraine, 2014. http://www.theses.fr/2014LORR0001/document.
Full textRecently, the use L-b-L assembly of natural polyelectrolytes has been suggested as a new technique of surface modification. The aim of our work was to develop multilayered films based on CHI and HA polymers bearing chemical. WJ-MSCs were seeded on these substrates. Biocompatibility was assessed by SEM and actin visualization. WJ-MSCs differentiation into endothelial phenotype was followed by western blot. First of all, SEM showed a fibroblastic morphology of adherent cells on CHI/HA and CHI-SH/HA-Thio as for control. WB analysis showed a rise of CD31, VEGF-R2 and VE-cadherin expression on CHI-SH/HA-Thio and CHI/HA. Films CHI-SH/HA-Thio were biocompatible and promoted endothelial differentiation potential