Structured PDMS Chambers for Enhanced Human Neuronal Cell Activity on MEA Platforms

Kreutzer, Joose; Ylä‐Outinen, Laura; Kärnä, Paula; Kaarela, Tiina; Mikkonen, Jarno E.; Skottman, Heli; Narkilahti, Susanna; Kallio, Pasi

doi:10.1016/s1672-6529(11)60091-7

Cited by 25 publications

(23 citation statements)

References 41 publications

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“…The 1-well MEA control is an open volume system with no liquid volume restrictions. To analyse the development of network activity, the data were compared to recordings from 60-6wellMEA200/30iR-Ti MEAs (MCS), in which the individual wells were separated using SpikeBooster devices (BioMediTech) (Kreutzer et al, 2012 ), and is referred to as the 6-well MEA control. The dimensions of the cell culturing areas on the SpikeBooster devices are the same as the cell plating areas on the tunnel devices, and the 6-well MEA control can be considered a partially restricted volume system.…”

Section: Methodsmentioning

confidence: 99%

“…Microengineered polydimethylsiloxane (PDMS) devices can be used to answer specific questions on mechanisms of neural function and pathology (Taylor et al, 2005 ; Scott et al, 2013 ; Ren et al, 2015 ), and they can also be used to facilitate the analysis of electrical function of hPSC-derived networks. PDMS devices consisting an open chamber (or “well”) can guide the network to grow more densely on top of the measuring electrodes, and thus facilitate the development and detection of neuronal activity (Kreutzer et al, 2012 ). PDMS microtunnel devices, on the other hand, increase the detected activity by amplifying extracellular electrical signals detected by the MEA (FitzGerald et al, 2009 ; Wieringa et al, 2010 ; Wang et al, 2012 ).…”

Section: Introductionmentioning

confidence: 99%

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Optimised PDMS Tunnel Devices on MEAs Increase the Probability of Detecting Electrical Activity from Human Stem Cell-Derived Neuronal Networks

et al. 2017

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Measurement of the activity of human pluripotent stem cell (hPSC)-derived neuronal networks with microelectrode arrays (MEAs) plays an important role in functional in vitro brain modelling and in neurotoxicological screening. The previously reported hPSC-derived neuronal networks do not, however, exhibit repeatable, stable functional network characteristics similar to rodent cortical cultures, making the interpretation of results difficult. In earlier studies, microtunnels have been used both to control and guide cell growth and amplify the axonal signals of rodent neurons. The aim of the current study was to develop tunnel devices that would facilitate signalling and/or signal detection in entire hPSC-derived neuronal networks containing not only axons, but also somata and dendrites. Therefore, MEA-compatible polydimethylsiloxane (PDMS) tunnel devices with 8 different dimensions were created. The hPSC-derived neurons were cultured in the tunnel devices on MEAs, and the spontaneous electrical activity of the networks was measured for 5 weeks. Although the tunnel devices improved the signal-to-noise ratio only by 1.3-fold at best, they significantly increased the percentage of electrodes detecting neuronal activity (52–100%) compared with the controls (27%). Significantly higher spike and burst counts were also obtained using the tunnel devices. Neuronal networks inside the tunnels were amenable to pharmacological manipulation. The results suggest that tunnel devices encompassing the entire neuronal network can increase the measured spontaneous activity in hPSC-derived neuronal networks on MEAs. Therefore, they can increase the efficiency of functional studies of hPSC-derived networks on MEAs.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimised PDMS Tunnel Devices on MEAs Increase the Probability of Detecting Electrical Activity from Human Stem Cell-Derived Neuronal Networks

et al. 2017

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“…To analyse the neuronal activity, a single cell suspension was also replated in NDM on 0.05% polyethyleneimine and 20 µg/ml human laminin coated MEA plates ( n = 8, 6wellMEA200/30iR-Ti, Multi Channel Systems, Germany) containing custom-made poly(dimethylsiloxane, PDMS) culturing chambers (Kreutzer et al, 2012). These chambers have capacity of 200 µl of medium/well and the cell attachment area is limited to 7 mm 2 .…”

Section: Methodsmentioning

confidence: 99%

Healthy human CSF promotes glial differentiation of hESC-derived neural cells while retaining spontaneous activity in existing neuronal networks

et al. 2013

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SummaryThe possibilities of human pluripotent stem cell-derived neural cells from the basic research tool to a treatment option in regenerative medicine have been well recognized. These cells also offer an interesting tool for in vitro models of neuronal networks to be used for drug screening and neurotoxicological studies and for patient/disease specific in vitro models. Here, as aiming to develop a reductionistic in vitro human neuronal network model, we tested whether human embryonic stem cell (hESC)-derived neural cells could be cultured in human cerebrospinal fluid (CSF) in order to better mimic the in vivo conditions. Our results showed that CSF altered the differentiation of hESC-derived neural cells towards glial cells at the expense of neuronal differentiation. The proliferation rate was reduced in CSF cultures. However, even though the use of CSF as the culture medium altered the glial vs. neuronal differentiation rate, the pre-existing spontaneous activity of the neuronal networks persisted throughout the study. These results suggest that it is possible to develop fully human cell and culture-based environments that can further be modified for various in vitro modeling purposes.

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“…PSCs can be encapsulated in structural materials such as ECM [75] or in spatially organized, 3D hydrogels that incorporated signaling factors [76]. Microwells have even been developed in conjunction with microelectrode arrays that can be used to characterize electrical activity of cells [77]. Microwells have even been designed so that the bottom face is a fabricated electrospun network of nano/microfibers [78].…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Microwell Regulation of Pluripotent Stem Cell Self-Renewal and Differentiation

Hsiao

Palecek

2012

BioNanoSci.

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The fates of pluripotent stem cells (PSCs), including survival, self-renewal, and differentiation, are regulated by chemical and mechanical cues presented in the three-dimensional (3D) microenvironment. Most PSC studies have been performed on two-dimensional substrates. However, 3D culture systems have demonstrated the importance of intercellular interactions in regulating PSC self-renewal and differentiation. Microwell culture systems have been developed to generate homogenous PSC colonies of defined sizes and shapes and to study how colony morphology affects cell fate. Using microwells, researchers have demonstrated that PSCs remain in a self-renewing undifferentiated state as a result of autocrine and paracrine signaling. Other studies have shown that microwell regulation of embryoid body size affects lineage commitment during differentiation via cell-cell contact and expression of soluble signals. In this review, we discuss recent advances in the design and utilization of 3D microwell platforms for studying intercellular regulation of PSC cell fate decisions and the underlying molecular mechanisms.

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Structured PDMS Chambers for Enhanced Human Neuronal Cell Activity on MEA Platforms

Cited by 25 publications

References 41 publications

Optimised PDMS Tunnel Devices on MEAs Increase the Probability of Detecting Electrical Activity from Human Stem Cell-Derived Neuronal Networks

Optimised PDMS Tunnel Devices on MEAs Increase the Probability of Detecting Electrical Activity from Human Stem Cell-Derived Neuronal Networks

Healthy human CSF promotes glial differentiation of hESC-derived neural cells while retaining spontaneous activity in existing neuronal networks

Microwell Regulation of Pluripotent Stem Cell Self-Renewal and Differentiation

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