<div class="csl-bib-body">
<div class="csl-entry">Remmers, D. (2015). <i>Influences of varying oxygen, glucose, and lactate concentrations on the osteogenic development of adipose derived mesenchymal stem cells</i> [Diploma Thesis, Technische Universität Wien]. reposiTUm. https://doi.org/10.34726/hss.2015.23150</div>
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dc.identifier.uri
https://doi.org/10.34726/hss.2015.23150
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/6242
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dc.description.abstract
The aim of this work is to investigate the impact of varying cell culture conditions on the osteogenic differentiation of adMSCs. Glucose and oxygen supply are two parameters which are known for their strong impact on cell behavior. Therefore, the effect of glucose and atmospheric oxygen concentration are examined in this study individually and in combination. Cells are cultivated over a period of 12 days under either high (21%) or low (5%) atmospheric oxygen concentrations and in osteogenic differentiation media with high (4%) or low (1%) glucose concentration. Medium change is carried out in a batch-process approach: A complete medium change is performed every three days to replenish glucose as well as other nutrients and growth factors and to remove lactate and other waste products. Daily measurements of glucose and lactate are performed to compare metabolic activity of the cells under the different cultivation conditions. Special focus is put on discriminating between the effects of glucose concentration and the metabolic changes in response to rising lactate levels. Differentiation is monitored by staining for ECM mineralization: Alizarin red and calcein staining are performed to detect calcification, while phosphate deposition is visualized by von Kossa staining. Cell load and cell distribution are shown by DAPI fluorescence staining of cell nuclei. Results of the study show glucose restriction in the presence of the low- glucose medium. Based on staining results for normoxia experiencing adMSCs, low- and high-glucose media lead to comparable results concerning the osteogenic differentiation. Regarding oxygen availability normoxic (21% oxygen) conditions result in lower glucose consumption rates and a stronger and earlier ECM ossification compared to atmospheric conditions of 5% oxygen. Alizarin red staining significantly detects ECM calcification as early as on differentiation day 7 when cultured under normoxic conditions. This behavior is observed for young (passage zero) adMSCs and passage number of the donor cells could be a crucial impact on the cell response to normoxic and hypoxic conditions. Generally, earlier and stronger mineralization is observed close to the border of the cell culture vessel. The study also shows a correlation between low lactate yield ratios and strong osteogenic differentiation.
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dc.language
English
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dc.language.iso
en
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dc.rights.uri
http://rightsstatements.org/vocab/InC/1.0/
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dc.subject
Stammzellen
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dc.subject
mesenchymale Stammzellen
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dc.subject
multipotente Stromazellen
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dc.subject
Zucker
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dc.subject
Laktat
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dc.subject
Laktatproduktion
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dc.subject
Glukosekonzentration
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dc.subject
Laktatkonzentration
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dc.subject
mechanische Stimuli
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dc.subject
Tissue Engineering
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dc.subject
Hypoxie
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dc.subject
Sauerstoff
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dc.subject
Sauerstoffkonzentration
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dc.subject
Respiration
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dc.subject
lactate yield coefficient
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dc.subject
Differenzierung
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dc.subject
Osteogenese
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dc.subject
Mechanosensation
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dc.subject
tägliche Messung
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dc.subject
optimale Bedingungen
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dc.subject
Luftzusammensetzung
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täglich
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dc.subject
Laktatakkumulation
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dc.subject
¿MEM
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dc.subject
DMEM
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dc.subject
stem cells
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dc.subject
mesenchymal stem cells
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dc.subject
multipotent stromal cells
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dc.subject
sugar
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dc.subject
lactate
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dc.subject
lactate production
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dc.subject
sugar supply
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dc.subject
glucose concentration
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dc.subject
lactate supply
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dc.subject
mechanical stimuli
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dc.subject
tissue engineering
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dc.subject
hypoxia
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dc.subject
oxygen
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dc.subject
oxygen concentration
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dc.subject
respiration
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dc.subject
lactate yield coefficient
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dc.subject
differentiation
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dc.subject
osteogenic
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dc.subject
mechanosensation
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dc.subject
osteogenesis
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dc.subject
daily examination
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dc.subject
optimal conditions
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dc.subject
atmospheric comparison
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dc.subject
daily
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dc.subject
lactate accumulation
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dc.subject
¿MEM
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dc.subject
DMEM
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dc.subject
time charts
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dc.title
Influences of varying oxygen, glucose, and lactate concentrations on the osteogenic development of adipose derived mesenchymal stem cells
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dc.type
Thesis
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dc.type
Hochschulschrift
de
dc.rights.license
In Copyright
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dc.rights.license
Urheberrechtsschutz
de
dc.identifier.doi
10.34726/hss.2015.23150
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dc.contributor.affiliation
TU Wien, Österreich
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dc.rights.holder
Daniel Remmers
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tuw.version
vor
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tuw.thesisinformation
Technische Universität Wien
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tuw.publication.orgunit
E300 - Fakultät für Maschinenwesen und Betriebswissenschaften