High throughput assembly of spatially controlled 3D cell clusters on a micro/nanoplatform

Abstract

Guided assembly of microscale tissue subunits (i.e. 3D cell clusters/aggregates) has found applications in cell therapy/tissue engineering, cell and developmental biology, and drug discovery. As cluster size and geometry are known to influence cellular responses, the ability to spatially control cluster formation in a high throughput manner could be advantageous for many biomedical applications. In this work, a micro- and nanofabricated platform was developed for this purpose, consisting of a soft-lithographically fabricated array of through-thickness microwells structurally bonded to a sheet of electrospun fibers. The microwells and fibers were manufactured from several polymers of biomedical interest. Human hepatocytes were used as model cells to demonstrate the ability of the platform to allow controlled cluster formation. In addition, the ability of the device to support studies on semi-controlled heterotypic interactions was demonstrated by co-culturing hepatocytes and fibroblasts. Preliminary experiments with other cells of interest (pancreatic cells, embryonic stem cells, and cardiomyocytes) were also conducted. Our platform possesses several advantages over previously developed microwell arrays: a more in vivo-like topographical stimulation of cells; better nutrient/waste exchange through the underlying nanofiber mat; and easy integration into standard two-chamber cell culture well systems.

Transwells with Microstamped Membranes Produce Micropatterned Two-Dimensional and Three-Dimensional Co-Cultures

“…[24][25][26][27][28][29][30][31][32] For example, microwell systems have been utilized for ES cell studies to control the formation of ES cellular aggregates because the size of aggregates regulates ES cellular differentiation. [33][34][35] Here, we present a simple and rapid cell patterning method to provide versatile control over the geometry of cellular niches formed using readily available Transwells.…”

PPO/PEO modified hollow fiber membranes improved sensitivity of 3D cultured hepatocytes to drug toxicity via suppressing drug adsorption on membranes

“…This study indicated that the resistance of drug adsorption could only be obtained by simultaneously increasing hydrophobicity and reducing electrostatic interaction. It is quite different from the protein resistance via simply utilizing the hydrophilic interaction [5,20,21]. The hydrophilic PEO modification successfully reduced protein adsorption to less than 10% of that on unmodified surfaces [5,20] but only had a little effect on reducing base drug adsorption.…”

“…It is quite different from the protein resistance via simply utilizing the hydrophilic interaction [5,20,21]. The hydrophilic PEO modification successfully reduced protein adsorption to less than 10% of that on unmodified surfaces [5,20] but only had a little effect on reducing base drug adsorption. Reducing the drug adsorption is more difficult than protein adsorption, because most of the drugs are positively charged which readily adsorb onto negatively charged synthetic polymers [2][3][4] while the proteins are mostly negatively charged at pH 7.4 that could avoid this problem [22].…”

Microwell perfusion array for high-throughput, long-term imaging of clonal growth

“…4,5 Constraining cell movement is necessary for atomic force microscopy 6 or to study the formation of cell clusters. 7,8 Lecault and colleagues 9 employed 160 lm cubic microchambers to study clonal culture of primitive mouse hematopoietic stem cells using live-cell imaging. 9 Development of microdevices for long-term perfusion culture is a significant bioengineering challenge requiring in-depth analysis and optimisation of system properties.…”