The negative photoresist SU-8 has found widespread use as a material in the fabrication of microelectrical-mechanical systems (MEMS). While SU-8 has been utilized as a structural material for biological MEMS, a number of SU-8 properties limit its application in these bioanalytical devices. These attributes include its brittleness, nonspecific adsorption of biomolecules, and high fluorescence in the visible wavelengths. In addition, native SU-8 is a poor substrate for cellular adhesion. Photoresists composed of resins with epoxide side groups and photoacids were screened for their ability to serve as a low fluorescence photoresist with sufficient resolution to generate microstructures with dimensions of 5-10 μm. The fluorescence of structures formed from 1002F photoresist (1002F resin combined with triarylsulfonium hexafluoroantimonate salts) was as much as 10 times less fluorescent than similar SU-8 microstructures. The absorbance of 1002F in the visible wavelengths was also substantially lower than that of SU-8. Microstructures or pallets with an aspect ratio as high as 4:1 could be formed permitting 1002F to be used as a structural material in the fabrication of arrays of pallets for sorting adherent cells. Several different cell types were able to adhere to native 1002F surfaces and the viability of these cells was excellent. As with SU-8, 1002F has a weak adhesion to glass, a favorable attribute when the pallet arrays are used to sort adherent cells. A threshold, laserpulse energy of 3.5 μJ was required to release individual 50-μm, 1002F pallets from an array. Relative to SU-8, 1002F photoresist offers substantial improvements as a substrate in bioanalytical devices and is likely to find widespread use in BioMEMS.
Antibodies are an important component in host immune responses to viral pathogens. Because of their unique maturation process, antibodies can evolve to be highly specific to viral antigens. Physicians and researchers have been relying on such high specificity in their quest to understand host–viral interaction and viral pathogenesis mechanisms and to find potential cures for viral infection and disease. With more than 60 recombinant monoclonal antibodies developed for human use in the last 20 years, monoclonal antibodies are now considered a viable therapeutic modality for infectious disease targets, including newly emerging viral pathogens such as Ebola representing heightened public health concerns, as well as pathogens that have long been known, such as human cytomegalovirus. Here, we summarize some recent advances in identification and characterization of monoclonal antibodies suitable as drug candidates for clinical evaluation, and review some promising candidates in the development pipeline.
The selection and collection of single cells from within a heterogeneous population is required to produce genetically engineered cell lines, to develop new stem cell lines, and for single-cell studies. We describe a new platform for the positive selection of single live mammalian cells while the cells remain adherent to their growth surface. Cells were grown on arrays of microfabricated, releasable elements composed of SU-8 polymer termed "cell pallets". The presence of air between the elements restricted the cells to the top surfaces of the pallets. Single pallets situated within large arrays of pallets were released on demand using a single, focused, laser pulse. The laser pulses were low in energy (2-5 muJ) and did not detach nearby, nontargeted pallets. Since the SU-8 pallets and the underlying glass substrate were optically transparent, the cells on the pallets could be visualized by microscopy before and after release. Over 90% of cells remained attached to the pallet during laser-based release. The feasibility of growing the cells from the released pallets into clonal colonies was demonstrated. The pallet array system permits adherent cells to be inspected using conventional microscopy and selected cells released for further analysis. The ability to assess cells while they remain adherent to a surface will broaden the number of attributes that can be utilized for cell separation, for example, cell shape, cytoskeletal properties, and other attributes.
Mesenchymal stem cells (MSCs) possess self-renewal and multipotential differentiation abilities, and they are thought to be one of the most reliable stem cell sources for a variety of cell therapies. Recently, cell therapy using MSCs has been studied as a novel therapeutic approach for cancers that show refractory progress and poor prognosis. MSCs from different tissues have different properties. However, the effect of different MSC properties on their application in anticancer therapies has not been thoroughly investigated. In this study, to characterize the anticancer therapeutic application of MSCs from different sources, we established two different kinds of human MSCs: umbilical cord blood-derived MSCs (UCB-MSCs) and adipose-tissue-derived MSCs (AT-MSCs). We used these MSCs in a coculture assay with primary glioblastoma multiforme (GBM) cells to analyze how MSCs from different sources can inhibit GBM growth. We found that UCB-MSCs inhibited GBM growth and caused apoptosis, but AT-MSCs promoted GBM growth. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick-end labeling assay clearly demonstrated that UCB-MSCs promoted apoptosis of GBM via tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). TRAIL was expressed more highly by UCBMSCs than by AT-MSCs. Higher mRNA expression levels of angiogenic factors (vascular endothelial growth factor, angiopoietin 1, platelet-derived growth factor, and insulin-like growth factor) and stromal-derived factor-1 (SDF-1/CXCL12) were observed in AT-MSCs, and highly vascularized tumors were developed when ATMSCs and GBM were cotransplanted. Importantly, CXCL12 inhibited TRAIL activation of the apoptotic pathway in GBM, suggesting that AT-MSCs may support GBM development in vivo by at least two distinct mechanisms-promoting angiogenesis and inhibiting apoptosis. The opposite effects of AT-MSCs and UCBMSCs on GBM clearly demonstrate that differences must be considered when choosing a stem cell source for safety in clinical application.
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