The fabrication of cell-laden structures with anisotropic mechanical
properties while having a precise control over the distribution of different
cell types within the constructs is important for many tissue engineering
applications. Automated textile technologies for making fabrics allow
simultaneous control over the color pattern and directional mechanical
properties. The use of textile techniques in tissue engineering, however,
demands the presence of cell-laden fibers that can withstand the mechanical
stresses during the assembly process. Here, the concept of composite living
fibers (CLFs) in which a core of load bearing synthetic polymer is coated by a
hydrogel layer containing cells or microparticles is introduced. The core thread
is drawn sequentially through reservoirs containing a cell-laden prepolymer and
a crosslinking reagent. The thickness of the hydrogel layer increases linearly
with to the drawing speed and the prepolymer viscosity. CLFs are fabricated and
assembled using regular textile processes including weaving, knitting, braiding,
winding, and embroidering, to form cell-laden structures. Cellular viability and
metabolic activity are preserved during CLF fabrication and assembly,
demonstrating the feasibility of using these processes for engineering
functional 3D tissue constructs.
DNA microarrays were rapidly scaled up from 256 to 6.5 million targets, and although antibody microarrays were proposed earlier, sensitive multiplex sandwich assays have only been scaled up to a few tens of targets. Cross-reactivity, arising because detection antibodies are mixed, is a known weakness of multiplex sandwich assays that is mitigated by lengthy optimization. Here, we introduce (1) vulnerability as a metric for assays. The vulnerability of multiplex sandwich assays to cross-reactivity increases quadratically with the number of targets, and together with experimental results, substantiates that scaling up of multiplex sandwich assays is unfeasible. We propose (2) a novel concept for multiplexing without mixing named antibody colocalization microarray (ACM). In ACMs, both capture and detection antibodies are physically colocalized by spotting to the same two-dimensional coordinate. Following spotting of the capture antibodies, the chip is removed from the arrayer, incubated with the sample, placed back onto the arrayer and then spotted with the detection antibodies. ACMs with up to 50 targets were produced, along with a binding curve for each protein. The ACM was validated by comparing it to ELISA and to a small-scale, conventional multiplex sandwich assay (MSA). Using ACMs, proteins in the serum of breast cancer patients and healthy controls were quantified, and six candidate biomarkers identified. Our results indicate that ACMs are sensitive, robust, and scalable.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.