You are hereAugust 6, 2015 | NSCs/NPCs
3D Neuron Growth Strategy to Boost the Study of Parkinson’s Disease
Review of “Differentiation of neuroepithelial stem cells into functional dopaminergic neurons in 3D microfluidic cell culture” from Lab Chip by Stuart P. Atkinson
Cinematography, printing, and cell culture seem at first glance to be distinct unrelated concepts, although all have recently embraced the move to from 2D to the more realistic 3D. This move has been especially important and successful in cell growth and differentiation, where 3D growth allows the recapitulation of the in vivo environment. Jens C. Schwamborn and Ronan M. T. Fleming from the University of Luxembourg have recently reported on their success with differentiating dopaminergic neurons (DNs) from human neuroepithelial stem cells (hNESCs) derived from induced pluripotent stem cells (iPSCs)  in microfluidic cell culture. This allows precise control over the cellular microenvironment, enhanced monitoring, and controlled perfusion culture [2-4], and the researchers hope that strategy will represent an economic and realistic means to produce a model system for the study of disorders such as Parkinson’s Disease .
The research used the OrganoPlate microfluidic cell culture platform , in which miniature bioreactors are fed by wells from a 365 well plate to allow phase-guided 3D cell growth. Briefly, one well is used to load cell-infused Matrigel growth substrate creating a culture lane, while media flows in a perfusion lane alongside (See the great video describing the system here). 2-lane OrganoPlates allow for simple “one-layered” growth while 3-lane OrganoPlates mediate stratified growth conditions (multiple gels or multiple media perfusion) if required.
Following their generation from iPSCs, the researchers introduced hNESCs into the OrganoPlates culture lane where they formed aggregates and began to differentiate and mature into dopaminergic neurons over a 30 day period using a modification of a previously published protocol . In the 2-lane plates, the authors found 72% of cells positive for the neuron marker TUBβIII and 19% double positive for TUBβIII and the DN marker TH, while the 3-lane plates which used two medium perfusion channels surrounding the cell-laden gel gave slightly lower percentages. This indicates that the OrganoPlate microfluidic cell culture platform permitted efficient differentiation into neurons expressing dopaminergic markers. One further advantage of this culture system is the ability to easily assess the 3D distribution of differentiated cells, and in this study, this permitted the researchers to see high levels of branching and cell interconnectivity, and to assess the length of neurites of differentiated neurons. The group finally tested the functionality of differentiated neurons and demonstrated spontaneous calcium transients evoked by action potentials in TH-positive cells consistent with previous in vitro studies on the electrophysiological activity of dopaminergic neurons.
This confirms the ability and biocompatibility of this new system to differentiate stem cells into specific cell types, and brings with it multiple advantages. These include the steep decrease in media and growth substrate requirements, and the ease of computerized automation of the entire process, which may encourage industrial-scale production of cells for both laboratory study, and even clinical-grade cell production strategies.
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