Bioengineering / Biyomühendislik
Permanent URI for this collectionhttps://hdl.handle.net/11147/4529
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Conference Object Deep Convolutional Neural Networks for Viability Analysis Directly From Cell Holograms Captured Using Lensless Holographic Microscopy(The Chemical and Biological Microsystems Society (CBMS), 2019) Delikoyun, Kerem; Çine, Ersin; Anıl İnevi, Müge; Özçivici, Engin; Özuysal, Mustafa; Tekin, Hüseyin CumhurCell viability analysis is one of the most widely used protocols in the fields of biomedical sciences. Traditional methods are prone to human error and require high-cost and bulky instrumentations. Lensless digital inline holographic microscopy (LDIHM) offers low-cost and high resolution imaging. However, recorded holograms should be digitally reconstructed to obtain real images, which requires intense computational work. We introduce a deep transfer learning-based cell viability classification method that directly processes the hologram without reconstruction. This new model is only trained once and viability of each cell can be predicted from its hologram. © 2019 CBMS-0001.Article Citation - WoS: 79Citation - Scopus: 93Magnetic Force-Based Micro Fluidic Techniques for Cellular and Tissue Bioengineering(Frontiers Media S.A., 2018) Yaman, Sena; Anıl İnevi, Müge; Özçivici, Engin; Tekin, Hüseyin CumhurLive cell manipulation is an important biotechnological tool for cellular and tissue level bioengineering applications due to its capacity for guiding cells for separation, isolation, concentration, and patterning. Magnetic force-based cell manipulation methods offer several advantages, such as low adverse effects on cell viability and low interference with the cellular environment. Furthermore, magnetic-based operations can be readily combined with microfluidic principles by precisely allowing control over the spatiotemporal distribution of physical and chemical factors for cell manipulation. In this review, we present recent applications of magnetic force-based cell manipulation in cellular and tissue bioengineering with an emphasis on applications with microfluidic components. Following an introduction of the theoretical background of magnetic manipulation, components of magnetic force-based cell manipulation systems are described. Thereafter, different applications, including separation of certain cell fractions, enrichment of rare cells, and guidance of cells into specific macro- or micro-arrangements to mimic natural cell organization and function, are explained. Finally, we discuss the current challenges and limitations of magnetic cell manipulation technologies in microfluidic devices with an outlook on future developments in the field.Conference Object Citation - WoS: 5Citation - Scopus: 8Lensless Digital In-Line Holographic Microscopy for Space Biotechnology Applications(Institute of Electrical and Electronics Engineers Inc., 2019) Delikoyun, Kerem; Çine, Ersin; Anıl İnevi, Müge; Özuysal, Mustafa; Özçivici, Engin; Tekin, Hüseyin CumhurBiomechanical changes at cellular level can dramatically affect living organisms in both aviation and space applications. Weightlessness induces morphological alteration of cells, which leads to tissue loss. Therefore, scientists have been studying the effect of weightlessness using cell culture based biological experiments using conventional microscopes. However, strict requirements regarding cost, weight and functionality limit the use of conventional microscopes in space environment. Lensless digital in-line holographic microscopy enables to use low-weight, low-cost and robust elements, such as a light emitting diode (LED), an aperture and an imaging sensor, instead of bulky, expensive and fragile optical elements, such as lenses, mirrors and filters. This technology offers a high field of view compared to conventional microscopes without affecting the resolution and it is also suitable for remote sensing applications with automated imaging capabilities. Here, we present a portable digital in-line holographic microscopy platform that allows to visualize cells and to analyze their viability in a microfluidic chip. The platform offers microscopic imaging with 1.55 mu m spatial resolution, 21.7 mm(2) field of view and image coloring capability. This platform could potentially play an important role in space biotechnology applications by enabling low-cost, high-resolution and portable monitoring of cells.Conference Object Citation - WoS: 7Citation - Scopus: 7Cell Separation With Hybrid Magnetic Levitation-Based Lensless Holographic Microscopy Platform(Institute of Electrical and Electronics Engineers Inc., 2019) Delikoyun, Kerem; Yaman, Sena; Anıl İnevi, Müge; Özçivici, Engin; Tekin, Hüseyin CumhurSeparation of target cells in a heterogeneous solution is of great importance for clinical studies especially for immunology and oncology. Separated cells can be used for diagnostic applications ranging from whole blood counting to isolation of circulating tumor cells (CTC) for personalized medicine. Recent separation technologies rely on labelling and identifying target cells with variety of labelling principle such as fluorescence or magnetic tags. However, they require labor-intensive processes, long analysis time, and expensive chemical reagents and instrumentation. Hence, their usage is limited to well-equipped centralized laboratories. There is a need for a rapid, sensitive, low-cost and automated cell separation technology to disseminate usage of this technology even in rural areas. Magnetic levitation is a powerful cell separation method, which distinguishes cells based on their levitation heights depending on cell density. However, magnetic levitation-based separation technologies require traditional, bulky and expensive microscopes for analysis. Lensless digital inline holographic microscopy (LDIHM) systems are composed of a simple illumination system containing an LED, a pinhole, and an imaging sensor for high-resolution microscopic imaging, which eliminates needs of highly fragile and expensive optics as in traditional microscopy. Here, we introduced a novel hybrid and portable cell separation platform, where magnetic levitation technology is integrated with LDIHM system for automated analysis of cell levitation heights. Using this platform, three different cell lines are successfully separated. Live and dead cells having distinguished levitation heights can be also identified in the platform.Article Citation - WoS: 34Citation - Scopus: 43Label-Free Density-Based Detection of Adipocytes of Bone Marrow Origin Using Magnetic Levitation(Royal Society of Chemistry, 2019) Sarıgil, Öykü; Anıl İnevi, Müge; Yılmaz, Esra; Meşe, Gülistan; Tekin, Hüseyin Cumhur; Özçivici, EnginAdipocyte hypertrophy and hyperplasia are important parameters in describing abnormalities in adipogenesis that are concomitant to diseases such as obesity, diabetes, anorexia nervosa and osteoporosis. Therefore, technical developments in the detection of adipocytes become an important driving factor in adipogenesis research. Current techniques such as optical microscopy and flow cytometry are available in detection and examination of adipocytes, driving cell- and molecular-based research of adipogenesis. Even though microscopy techniques are common and straightforward, they are restricted in terms of manipulation and separation of the cells. Flow cytometry is an alternative, but mature adipocytes are fragile and cannot withstand the flow process. Other separation methods usually require labeling of the cells or usage of microfluidic platforms that utilize fluids with different densities. Magnetic levitation is a novel label-free technology with the principle of movement of cells towards the lower magnetic field in a paramagnetic medium depending on their individual densities. In this study, we used a magnetic levitation device for density-based single cell detection of differentiated adipogenic cells in heterogeneous populations. Results showed that the magnetic levitation platform was sensitive to changes in the lipid content of mesenchymal stem cells committed to adipogenesis and it could be successfully used to detect the adipogenic differentiation of the cells.Conference Object Citation - WoS: 3Citation - Scopus: 4Biofabrication of Cellular Structures Using Weightlessness as a Biotechnological Tool(IEEE, 2019) Anıl İnevi, Müge; Sarıgil, Öykü; Yaman, Sena; Yalçın Özuysal, Özden; Meşe, Gülistan; Tekin, Hüseyin Cumhur; Özçivici, EnginGravity is an important biomechanical signal effecting the morphology and function of organisms. Reduction of gravitational forces, as experienced during spaceflight, cause alterations in the biological systems. Magnetic levitation technique is one of the most recent ground-based technology to mimic weightlessness environment. In addition to providing a platform to investigate biological effects of the weightlessness, this platform presents a novel opportunity to biofabricate 3-dimensional (3D) structures in a scaffold-and nozzle-free fashion. In this study, various controllable self-assembled 3D living structures were fabricated via magnetic levitation technique. This strategy may offer an easy and cost-effective opportunity for a wide range of space biotechnology researches.Conference Object Citation - WoS: 4Citation - Scopus: 5Application of Magnetic Levitation Induced Weightlessness To Detect Cell Lineage(IEEE, 2019) Sarıgil, Öykü; Anıl İnevi, Müge; Yılmaz, Esra; Çağan, Melike; Meşe, Gülistan; Tekin, Hüseyin Cumhur; Özçivici, EnginIdentification and classification of bone marrow cells is an important step for molecular biology and therapeutic studies related to bone marrow disorders such as osteoporosis or obesity. In this study, we applied magnetic levitation technology to induce a weightlessness environment to detect adipocytes and osteoblasts based on their single cell density. This biotechnological method can be used for separation of heterogeneous populations such as bone marrow once adapted to a continuous microfluidic platform.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.Article Citation - WoS: 6Citation - Scopus: 7A Biodesign Approach To Obtain High Yields of Biosimilars by Anti-Apoptotic Cell Engineering: A Case Study To Increase the Production Yield of Anti-Tnf Alpha Producing Recombinant Cho Cells(Humana Press, 2018) Gülce İz, Sultan; Anıl İnevi, Müge; Sağlam Metiner, Pelin; Ayyıldız Tamiş, Duygu; Kisbet, NazlıRecent developments in medical biotechnology have facilitated to enhance the production of monoclonal antibodies (mAbs) and recombinant proteins in mammalian cells. Human mAbs for clinical applications have focused on three areas, particularly cancer, immunological disorders, and infectious diseases. Tumor necrosis factor alpha (TNF-α), which has both proinflammatory and immunoregulatory functions, is an important target in biopharmaceutical industry. In this study, a humanized anti-TNF-α mAb producing stable CHO cell line which produces a biosimilar of Humira (adalimumab) was used. Adalimumab is a fully human anti-TNF mAb among the top-selling mAb products in recent years as a biosimilar. Products from mammalian cell bioprocesses are a derivative of cell viability and metabolism, which is mainly disrupted by cell death in bioreactors. Thus, different strategies are used to increase the product yield. Suppression of apoptosis, also called anti-apoptotic cell engineering, is the most remarkable strategy to enhance lifetime of cells for a longer production period. In fact, using anti-apoptotic cell engineering as a BioDesign approach was inspired by nature; nature gives prolonged life span to some cells like stem cells, tumor cells, and memory B and T cells, and researchers have been using this strategy for different purposes. In this study, as a biomimicry approach, anti-apoptotic cell engineering was used to increase the anti-TNF-α mAb production from the humanized anti-TNF-α mAb producing stable CHO cell line by Bcl-xL anti-apoptotic protein. It was shown that transient transfection of CHO cells by the Bcl-xL anti-apoptotic protein expressing plasmid prolonged the cell survival rate and protected cells from apoptosis. The transient expression of Bcl-xL using CHO cells enhanced the anti-TNF-α production. The production of anti-TNF-α in CHO cells was increased up to 215 mg/L with an increase of 160% after cells were transfected with Bcl-xL expressing plasmid with polyethylenimine (PEI) reagent at the ratio of 1:6 (DNA:PEI). In conclusion, the anti-apoptotic efficacy of the Bcl-xL expressing plasmid in humanized anti-TNF-α MAb producing stable CHO cells is compatible with curative effect for high efficiency recombinant protein production. Thus, this model can be used for large-scale production of biosimilars through transient Bcl-xL gene expression as a cost-effective method.