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A Multiparameter Approach to Separation and Clonal Analysis of Mammalian CellsAmaya, Peter 25 August 2017 (has links)
No description available.
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The Utilization of Multipotent Mesenchymal Stromal Cell Transplantation to Improve Fascia RepairBown, Andre B. J. 19 September 2013 (has links)
No description available.
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Early Cellular Influence of BMP12, Compared to BMP2, on Equine Superficial Digital Flexor Tenocytes and Bone Marrow Derived Mesenchymal Stem Cells in VitroMurray, Shannon J. 03 September 2009 (has links)
No description available.
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Sandwich-like systems to engineer the cellular microenvironmentBallester Beltrán, José 20 March 2015 (has links)
Abstract
While most of the in vitro cultures are carried out on bi-dimensional (2D)
substrates, most of the in vivo extracellular matrices are threedimensional
(3D). Consequently cells behave differently on 2D substrates
as a way to self-adaptation to a non-physiological environment. This fact
has encouraged the development of more relevant culture conditions
seeking to provide more representative models for biomedicine (e.g.
cancer, drug discovery and tissue engineering) and further insights into
any dimension-dependent biological mechanism. Different 3D culture
systems have been established though their variability and complexity
hinder their standardisation in common cell culture procedures. So, this
thesis deals with the dimensionality issue in cell/material interactions and
introduces sandwich-like microenvironments as a versatile tool to study
cell behaviour. Cells cultured within this system use both dorsal and
ventral receptors to adhere and spread, undergoing important changes
with respect to the 2D cultures and approaching to 3D conditions.
Stimulation of dorsal receptors has been previously addressed by
overlaying a protein gel on cells already attached on a 2D surface. Here we
propose a sandwich-like system that consists of two 2D surfaces so that
wider spectra of conditions can be investigated by changing the nature of
the substrate (material, topography…) and the protein coatings of both
ventral and dorsal sides.
Since sandwich culture provides an altered cellular adhesion
compared to the traditional 2D substrates by the excitation of the dorsal
receptors, changes in the intracellular signalling are expected, which
might alter important processes such as proliferation, morphology,
migration and differentiation. Hence this thesis evaluates the effect of
different sandwich culture parameters in cell behaviour.
First, cell fate upon adhesion was evaluated in terms of
morphology, proliferation and adhesion. Different conditions were studied
such as materials with different properties or protein coatings (dorsal and
ventral substrates), as well as the effect of sandwiching cells just after
seeding or after been allowed to adhere to the ventral substrate.
Interesting results were obtained such as the relationship between the
ability of cells to reorganise the ECM with cell morphology, proliferation
and adhesion, similarly as observed in 3D hydrogels (degradable vs nondegradable
systems).
Then, cell migration within sandwich culture was studied by live
imaging of a wound healing assay. Results revealed the key effect of both
ventral and dorsal substrates in determining the migration rate as well as
the migration mode used by cells. Moreover cells within the sandwich
culture migrating in the wound healing assay adopted an elongated cell
morphology that resembled cells migrating in other 3D systems. Beyond
differences in cell morphology and migration, dorsal stimulation
promoted cell remodelling of the extra-cellular matrix (ECM) over simple
ventral receptor activation in traditional 2D cultures.
Finally the effect of sandwich culture on cell differentiation was
evaluated. First we showed an increase in C2C12 myogenic differentiation
when cultured within the sandwich system. This enhancement was shown
to be dorsal stimulation dependent and related to an alteration of the
signalling pathway and the growth factor release. To determine if
sandwich culture leads only to myogenic differentiation or whether it
allows differentiation to other lineages, 4 different human mesenchymal
stem cells (hMSCs) lines were cultured under the same conditions. Results
showed the same sandwich environment triggered different cell
differentiation. This points out the importance of the microenvironment
cell niche in vivo, which highly influence cell fate, and thus the need of
mimicking it properly in vitro.
Overall, sandwich-like microenvironments switch cell behaviour
towards 3D-like patterns, demonstrating the importance of this versatile,
simple and robust approach to mimic cell microenvironments in vivo. / Ballester Beltrán, J. (2014). Sandwich-like systems to engineer the cellular microenvironment [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/48166
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Human umbilical cord matrix mesenchymal stem cells suppress the growth of breast cancer by expression of tumor suppressor genesOhta, Naomi January 1900 (has links)
Master of Science / Department of Anatomy and Physiology / Masaaki Tamura / Previous studies have shown that both human and rat umbilical cord matrix mesenchymal stem cells (UCMSC) possess the ability to control the growth of breast carcinoma cells. Comparative analysis of two types of UCMSC suggest that rat UCMSC-dependent growth regulation is significantly stronger than that of human UCMSC. Accordingly, the present study was designed to clarify their different tumoricidal abilities by analyzing gene expression profiles in two types of UCMSC. Gene expression profiles were studied by microarray analysis using Illumina HumanRef-8-V2 and RatRef-12 BeadChip for the respective UCMSC. The gene expression profiles were compared to untreated naïve UCMSC and those co-cultured with species-matched breast carcinoma cells; human UCMSC vs. MDA-231 human carcinoma cells and rat UCMSC vs. Mat B III rat carcinoma cells. The following selection criteria were used for the screening of candidate genes associated with UCMSC-dependent tumoricidal ability; 1) gene expression difference should be at least 1.5 fold between naive UCMSC and those co-cultured with breast carcinoma cells; 2) they must encode secretory proteins and 3) cell growth regulation-related proteins. These analyses screened 17 common genes from human and rat UCMSC. The comparison between the two sets of gene expression profiles identified that two tumor suppressor genes, adipose-differentiation related protein (ADRP) and follistatin (FST), were specifically up-regulated in rat UCMSC, but down-regulated in human UCMSC when they were co-cultured with the corresponding species’ breast carcinoma cells. The suppression of either protein by the addition of a specific neutralizing antibody in co-culture of rat UCMSC with Mat B III cells significantly abrogated UCMSC ability to attenuate the growth of carcinoma cells. Over-expression of both genes by adenovirus vector in human UCMSC enhanced their
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ability to suppress the growth of MDA-231 cells. In the breast carcinoma lung metastasis model generated with MDA-231 cells, systemic treatment with FST-over-expressing human UCMSC significantly attenuated the tumor burden. These results suggest that both ADRP and FST may play important roles in exhibiting stronger tumoricidal ability in rat UCMSC than human UCMSC and imply that human UCMSC can be transformed into stronger tumoricidal cells by enhancing tumor suppressor gene expression.
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Differential Expression of Surface Markers in Mouse Bone Marrow Mesenchymal Stromal Cell Subpopulations with Distinct Lineage CommitmentAnastassiadis, Konstantinos, Rostovskaya, Maria 18 January 2016 (has links) (PDF)
Bone marrow mesenchymal stromal cells (BM MSCs) represent a heterogeneous population of progenitors with potential for generation of skeletal tissues. However the identity of BM MSC subpopulations is poorly defined mainly due to the absence of specific markers allowing in situ localization of those cells and isolation of pure cell types. Here, we aimed at characterization of surface markers in mouse BM MSCs and in their subsets with distinct differentiation potential. Using conditionally immortalized BM MSCs we performed a screening with 176 antibodies and high-throughput flow cytometry, and found 33 markers expressed in MSCs, and among them 3 were novel for MSCs and 13 have not been reported for MSCs from mice. Furthermore, we obtained clonally derived MSC subpopulations and identified bipotential progenitors capable for osteo- and adipogenic differentiation, as well as monopotential osteogenic and adipogenic clones, and thus confirmed heterogeneity of MSCs. We found that expression of CD200 was characteristic for the clones with osteogenic potential, whereas SSEA4 marked adipogenic progenitors lacking osteogenic capacity, and CD140a was expressed in adipogenic cells independently of their efficiency for osteogenesis. We confirmed our observations in cell sorting experiments and further investigated the expression of those markers during the course of differentiation. Thus, our findings provide to our knowledge the most comprehensive characterization of surface antigens expression in mouse BM MSCs to date, and suggest CD200, SSEA4 and CD140a as markers differentially expressed in distinct types of MSC progenitors.
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Human Wharton’s jelly cells-isolation and characterization in different growth conditionsSeshareddy, Kiran Babu January 1900 (has links)
Master of Science / Department of Anatomy and Physiology / Mark L. Weiss / Wharton's jelly is a non-controversial source of mesenchymal stromal cells. Isolation of the cells is non-invasive and painless. The cells have been shown to have a wide array of therapeutic applications. They have improved symptoms when transplanted in a variety of animal disease models, can be used in tissue engineering applications to grow living tissue ex vivo for transplantation, and can be used as drug delivery vehicles in cancer therapy. The cells have also been shown to be non-immunogenic and immune suppressive. This thesis focuses on optimizing isolation protocols, culture protocols, cryopreservation, and characterization of cells in different growth conditions.
Results from the experiments indicate that isolation of cells by enzyme digestion yields cells consistently, a freezing mixture containing 90% FBS and 10% DMSO confers maximum viability, and the expression of mesenchymal stromal cell consensus markers does not change with passage and cryopreservation. The results of the experiments also show that cells grow at a higher rate in 5% oxygen culture conditions compared to 21% oxygen culture conditions, serum does not have an effect on growth of the cells, serum and oxygen do not have effects on the expression of mesenchymal stromal cell consensus markers and the cells are stable without nuclear abnormalities when grown in 5% oxygen and serum free conditions for six passages after first establishing in serum conditions.
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Parakrine Beeinflussung der Genexpression in vitro von chondrogenen Zellen in der Osteoarthrose / Paracrine modulation of the gene-expression in vitro of chondrogenic cells in osteoarthritisMarks, Phillip 23 March 2016 (has links)
No description available.
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Identification of pathways in liver repair potentially targeted by secretory proteins from human mesenchymal stem cellsWinkler, Sandra, Hempel, Madlen, Brückner, Sandra, Tautenhahn, Hans-Michael, Kaufmann, Roland, Christ, Bruno 19 July 2016 (has links) (PDF)
Background: The beneficial impact of mesenchymal stem cells (MSC) on both acute and chronic liver diseases has been confirmed, although the molecular mechanisms behind it remain elusive. We aim to identify factors secreted by undifferentiated and hepatocytic differentiated MSC
in vitro in order to delineate liver repair pathways potentially targeted by MSC. Methods: Secreted factors were determined by protein arrays and related pathways identified by biomathematical analyses. Results: MSC from adipose tissue and bone marrow expressed a similar pattern
of surface markers. After hepatocytic differentiation, CD54 (intercellular adhesion molecule 1, ICAM-1) increased and CD166 (activated leukocyte cell adhesion molecule, ALCAM) decreased. MSC secreted different factors before and after differentiation. These comprised cytokines involved in innate immunity and growth factors regulating liver regeneration. Pathway analysis revealed cytokine-cytokine receptor interactions, chemokine signalling pathways, the complement and coagulation cascades as well as the Januskinase-signal transducers and activators of transcription (JAK-STAT) and nucleotide-binding oligomerization domain-like receptor (NOD-like receptor) signalling pathways as relevant networks. Relationships to transforming growth factor beta(TGF-beta) and hypoxia-inducible factor 1-alpha (HIF1-alpha) signalling seemed also relevant. Conclusion: MSC secreted proteins, which differed depending on cell source and degree of differentiation. The factors might address inflammatory and growth factor pathways as well as chemo-attraction and innate immunity. Since these are prone to dysregulation in most liver diseases, MSC release hepatotropic factors, potentially supporting liver regeneration.
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Repair of skeletal muscle transection injury with tissue lossMerritt, Edward Kelly, 1979- 19 October 2009 (has links)
A traumatic skeletal muscle injury that involves the loss of a substantial portion of tissue will not regenerate on its own. Little is understood about the ability of the muscle to recover function after such a defect injury, and few research models exist to further elucidate the repair and regeneration processes of defected skeletal muscle. In the current research, a model of muscle injury was developed in the lateral gastrocnemius (LGAS) of the rat. In this model, the muscle gradually remodels but functional recovery does not occur over 42 days. Repair of the defect with muscle-derived extracellular matrix (ECM), improves the morphology of the LGAS. Blood vessels and myofibers grow into the ECM implant in vivo, but functional recovery does not occur. Addition of bone marrow-derived mesenchymal stem cells (MSCs) to the implanted ECM in the LGAS increases the number of blood vessels and regenerating myofibers within the ECM. Following 42 days of recovery, the cell-seeded ECM implanted LGAS produces significantly higher isometric force than the non-repaired and non-cell seeded ECM muscles. These results suggest that the LGAS muscle defect is a suitable model for the study of traumatic skeletal muscle injury with tissue loss. Additionally, MSCs seeded on an implanted ECM lead to functional restoration of the defected LGAS. / text
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