Monitoring the Differentiation and Migration Patterns of Neural Cells Derived from Human Embryonic Stem Cells Using a Microfluidic Culture System

Abstract

Microfluidics can provide unique experimental tools to visualize the development of neural structures within a microscale device, which is followed by guidance of neurite growth in the axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stem cells (hESCs). We co-cultured hESCs with PA6 stromal cells, and isolated neural rosette-like structures, which subsequently formed neurospheres in suspension culture. Tuj1-positive neural cells, but not nestin-positive neural precursor cells (NPCs), were able to enter the microfluidics grooves (microchannels), suggesting that neural cell-migratory capacity was dependent upon neuronal differentiation stage. We also showed that bundles of axons formed and extended into the microchannels. Taken together, these results demonstrated that microfluidics technology can provide useful tools to study neurite outgrowth and axon guidance of neural cells, which are derived from human embryonic stem cells.

Retrograde interferon‐gamma signaling induces major histocompatibility class I expression in human‐induced pluripotent stem cell‐derived neurons

“…In contrast, our chamber design and other recent designs provide an open access port that facilitates placement of the large neural aggregates near the microgrooves. 41,46 This design also facilitates media exchange without flow-induced disruption of the cells or shear force-induced damage to the axons.…”

“…MICs offer the ability to create growth factor or hormone gradients that mimic the in vivo microenvironment and the ability to compartmentalize extracellular matrix composition to enhance physical and functional differentiation. For neuron cultures in particular, MICs that separate axons from cell bodies have allowed scientists to study axon biology in greater detail, [40][41][42][43][44][45] permitting the analysis of factors that impact neurite outgrowth 46 and the analysis of retrograde and anterograde axonal signaling. 10 Building on these concepts, we provide a robust and efficient paradigm for transferring iPSC-derived, NSC-derived, neuron-rich neural aggregates into MICs that routinely achieves dense axonal outgrowth into distal chambers (Fig.…”

Microfluidic systems for stem cell-based neural tissue engineering

“…Combining diffusion and laminar flow, better controlled signaling, and the ability to co-culture cells in a 3D arrangement are the most important advantages of microfluidic cell culture 94 . Different applications of these characteristics in neural regeneration include the culture of single ESCs 95 , study of ESC differentiation and monitoring of their migration 96 , and gradient-mediated NSC chemotaxis 97 .…”

Full biological characterization of human pluripotent stem cells will open the door to translational research

“…Migration and invasion assays in stem cell research using hESC and hiPSC are routinely used to check proper cell differentiation into various cell types. Neurosphere migration assays (Topol et al 2015) and microfluidic culture systems (Lee et al 2014) were used to study neural cell-migratory capacity upon differentiation of hESC and hiPSC to neuronal cells, and scratch assays were used to validate migration potential of hiPSC-derived smooth muscle cells (Yang et al 2016). However, migration and invasion capacity of undifferentiated hESC and hiPSC has been poorly addressed so far.…”