bioRxiv Science⌕ Search

Biology subjects

Deshmukh, D.

Publications and source records attributed to Deshmukh, D..

2 recordsLinked to original sources

Acoustofluidic patterning for improved microtissue histology

Microtissues (such as organoids or spheroids) are useful to model healthy or diseased human tissues and organs, with application in drug testing, tissue engineering, and personalized medicine. To characterize their morphology and phenotype, microtissues are often monitored using histology and high-resolution imaging. While this technique is well-established and standard for macroscopic tissue sections, it remains challenging to execute and has low throughput in the context of these sub-millimeter structures, mainly due to the fact that their size makes them difficult to locate within the embedding medium. To enhance the efficiency of microtissue histology, we developed an acoustofluidic device that arranges microtissues in a coplanar arrangement within HistoGel, a widely used embedding medium. Pre-patterning microtissues within HistoGel streamlines downstream processing by positioning the centers of mass of the microtissues at a pre-defined plan in the sample, simplifying sectioning, labeling, and imaging. We validated the method using differently sized HepG2 microtissues and tumor microtissues from osteosarcoma cells (Saos-2, MG-63, HOS), demonstrating that this method can be applied to different microtissue shapes, sizes, and types, without modifying established post-processing techniques. Our approach substantially enhances the microtissue histology workflow and broadens possibilities for microtissue analysis.

bioengineering↗

Tunable bicontinuous macroporous cell culture scaffolds via kinetically controlled phase separation

Three-dimensional (3D) scaffolds enable biological investigations with a more natural cell conformation. However, the porosity of synthetic hydrogels is often limited to the nanometer scale, which confines the movement of 3D encapsulated cells and restricts dynamic cell processes. Precise control of hydrogel porosity across length scales remains a challenge and the development of porous materials that allow cell infiltration, spreading, and migration in a manner more similar to natural ECM environments is desirable. Here, we present a straightforward and reliable method for generating kinetically-controlled macroporous systems using liquid-liquid phase separation between poly(ethylene glycol) (PEG) and dextran. Photopolymerization-induced phase separation resulted in macroporous hydrogels with tunable pore size. Varying light intensity and hydrogel composition controlled polymerization kinetics, time to percolation, and complete gelation, which defined the average pore diameter (O = 1- 300 m) and final gel stiffness of the formed hydrogels. Critically, for biological applications, macroporous hydrogels were prepared from aqueous polymer solutions at physiological pH and temperature using visible light, allowing for direct cell encapsulation. We encapsulated human dermal fibroblasts in a range of macroporous gels with different pore sizes. Porosity improved cell spreading with respect to bulk gels and allowed migration in the porous systems.

bioengineering↗