Sürdürülebilir Yeşil Kampüs Koleksiyonu / Sustainable Green Campus Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7755
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Article Citation - WoS: 79Citation - Scopus: 94Biofabrication of in Situ Self Assembled 3d Cell Cultures in a Weightlessness Environment Generated Using Magnetic Levitation(Nature Publishing Group, 2018) Anıl İnevi, Müge; Yaman, Sena; Arslan Yıldız, Ahu; Meşe, Gülistan; Yalçın Özuysal, Özden; Tekin, Hüseyin Cumhur; Özçivici, EnginMagnetic levitation though negative magnetophoresis is a novel technology to simulate weightlessness and has recently found applications in material and biological sciences. Yet little is known about the ability of the magnetic levitation system to facilitate biofabrication of in situ three dimensional (3D) cellular structures. Here, we optimized a magnetic levitation though negative magnetophoresis protocol appropriate for long term levitated cell culture and developed an in situ 3D cellular assembly model with controlled cluster size and cellular pattern under simulated weightlessness. The developed strategy outlines a potential basis for the study of weightlessness on 3D living structures and with the opportunity for real-time imaging that is not possible with current ground-based simulated weightlessness techniques. The low-cost technique presented here may offer a wide range of biomedical applications in several research fields, including mechanobiology, drug discovery and developmental biology.Master Thesis Deciphering Functions of Aberrant Hemichannels Formed by Connexin 26- I30n and D50y Mutations(Izmir Institute of Technology, 2015) Aypek, Hande; Meşe Özçivici, GülistanCells need to communicate with each other for maintenance cellular and tissue homeostasis. Gap junctions are channel-forming structures that are formed by docking of two hemichannels on the plasma membrane of adjacent cells. Connexins are subunits of gap junctions. Connexin 26 (Cx26) is one of the connexin isoform and mutations on the Cx26 gene (GJB2) cause non-syndromic and syndromic deafness. Keratitis-ichthyosis-deafness (KID) syndrome is one of the syndromic deafness disorders caused by Cx26 mutations. Among these mutations, Cx26-I30N and D50Y missense mutations were shown to form aberrant hemichannels but their effect on protein biosynthesis and functions have not studied. In this study, we aimed to decipher in vitro functions of aberrant hemichannels formed by Cx26-I30N and D50Y mutations. First of all, the effect of Cx26-I30N and D50Y mutations on localization, mRNA expression and protein synthesis properties were investigated in HeLa, N2A and HaCaT cells. Results suggested that Cx26-I30N and D50Y mutants were not able to form gap junction plaques on the plasma membrane and were localized in the Golgi apparatus. In addition, mutations resulted in a reduction in mRNA expression and protein synthesis. After, functional analysis was performed in Cx26-I30N and D50Y transfected N2A and HaCaT cells. Internal Ca2+ content measurement, measurement of released ATP, measurement of cell size and apoptosis assays were performed. Ca2+ measurement results showed that both Cx26-I30N and D50Y mutations deregulate Ca2+ balance in both N2A and HaCaT cells. Result of ATP release assay indicated that ATP amount in the extracellular environment decreased in N2A cells having Cx26-I30N and D50Y clones. Finally, apoptosis assay showed that number of necrotic cells increased when N2A cells were transfected with Cx26-I30N and D50Y constructs. Therefore, it was shown that aberrant hemichannels formed by Cx26-I30N and D50Y mutations may induce necrotic cell death by disrupting Ca2+ balance and ATP amount in cells.Master Thesis Synthesis, Characterization of Cdsxse1-X Quantum Dots and Evaluation of Their Real-Time Motions in Live Cells(Izmir Institute of Technology, 2011) Ünal, Gülçin; Özçelik, SerdarThe use of quantum dots as fluorescent labels in bioimaging is the most intensively studied subject. The aim of this study is to elucidate locations of quantum dots and track their motions in real time through confocal microscopy and to evaluate influence of surface chemistry on diffusions of quantum dots in live cells. In this study, trioctylphosphine oxide (TOPO) capped CdSxSe1-x quantum dots were synthesized and then TOPO molecules were exchanged with 3-mercaptopropionic acid and N-acetyl-Lcysteine to make quantum dots water dispersible for cellular imaging. Human lung adenocarcinoma epithelial cells (A549) and human bronchial epithelial cells (BEAS-2B) were incubated 1 hour with CdSxSe1-x quantum dots with a concentration range of 1-10 g/mL. Localizations and real time motions of quantum dots were tracked by a spinning disc confocal microscope. The center of fluorescent spots of quantum dots was determined by 2D Gaussian fitting with a sub-pixel resolution (<100nm/pixel). The mean square displacements, diffusion coefficients and trajectories in which quantum dots made motions were analyzed by the software ImageJ with a plug in Spot Tracker. Confocal images showed that both MPA and NAC cappped quantum dots were observed in the cytoplasm of cells. Trajectories of quantum dots in cellular environment demonstrated that the quantum dots performed various types of motions in live cells. Unimodal, trimodal and multimodal distribution histograms of the diffusion coefficeints were obtained for different capping agents (MPA and NAC) and cell types (A549 and BEAS-2B). We conclude that the surface chemistry regulates the motion of the quantum dots in the cellular environment.