Compartmental culture of embryonic stem cell-derived neurons in microfluidic devices for use in axonal biology

Abstract

Axonal pathology has been clearly implicated in neurodegenerative diseases making the compartmental culture of neurons a useful research tool. Primary neurons have already been cultured in compartmental microfluidic devices but their derivation from an animal is a time-consuming and difficult work and has a limit in their sources. Embryonic stem cell (ESC)-derived neurons (ESC_Ns) overcome this limit, since ESCs can be renewed without limit and can be differentiated into ESC_Ns by robust and reproducible protocols. In this research, ESC_Ns were derived from mouse ESCs in compartmental microfluidic devices, and their axons were isolated from the somal cell bodies. Once embryoid bodies (EBs) were localized in the microfluidic culture chamber, ESC_Ns spread out from the EBs and occupied the cell culture chamber. Their axons traversed the microchannels and finally were isolated from the somata, providing an arrangement comparable to dissociated primary neurons. This ESC_N compartmental microfluidic culture system not only offers a substitute for the primary neuron counterpart system but also makes it possible to make comparisons between the two systems.

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

“…Furthermore, guidance of neurite growth, in the axonal isolation compartment in the device, can be monitored and recorded using time-lapse video microscopy (Hamon and Hong, 2013; Park et al, 2013). We have previously described a compartmental culture system of mouse embryonic stem cell (ESC)-derived neurons in microfluidic devices (Shin et al, 2010), and in the present report, have extended this approach to monitor the differentiation and migration of neural cells that are derived from hESCs. The results of the study demonstrated that microfluidics technology is a useful tool to study neurite outgrowth and axon guidance of neural cells that are derived from hESCs.…”

Microfluidic systems for stem cell-based neural tissue engineering

“…In some cases, especially with cell culture applications, charged molecules such as poly-D-lysine, or ECM proteins, including laminin, fibronectin and collagen, can be applied and coated onto the surface of PDMS 99 . Collagen type 1 115 , poly-L-lysine (PLL) 116 and fibronectin 111 coatings have all been used to overcome the shortcomings of PDMS, and other polymeric material-based microdevices. For example Pluronic F-127 treatment was used to prevent absorption of fluorescent molecules into the PDMS matrix, and lessening noise and background during fluorescence imaging 110 .…”

“…The ability to observe the different parts of neurons (axons and cell bodies) in compartmentalized microfluidic devices can be useful for studying neurodegenerative diseases. For example Hwa Sung Shin et al, used embryonic stem cell (ESC)-derived neurons (ESC_Ns) (derived from mouse ESCs) and showed that the axons traversed the microchannels, and were finally isolated from the somatic cell bodies to allow the study of axonal biology 116 (Fig. 4E).…”

“…E) Three-chamber microfluidic devices for Map5 stained axonal isolation of ESC_Ns (Reproduced from Ref. 116 . Copyright 2015 with permission from Springer).…”

Microfluidic devices for cell cultivation and proliferation

“…Shin et al 88 presented an axon-isolating microfluidic device for ESC-derived neurons to study neurodegenerative diseases such as axonal regeneration. The device consisted of two channels (containing holes) and joined by microgrooves.…”