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Dual-compartment neurofluidic system for electrophysiological measurements in physically segregated and functionally connected neuronal cell culture.

Kanagasabapathi TT, Ciliberti D, Martinoia S, Wadman WJ, Decré MM - Front Neuroeng (2011)

Bottom Line: Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated.The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining.In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques.

View Article: PubMed Central - PubMed

Affiliation: Minimally Invasive Healthcare Department, Philips Research Laboratories Eindhoven, Netherlands.

ABSTRACT
We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels.

No MeSH data available.


Related in: MedlinePlus

Structural connectivity between the two compartmentalized neuronal sub-populations. (A) Phase contrast image of neurite ladder structure intact after the removal of PDMS structures from the MEA surface; (B) Transfection image of a neurite grown across the microchannels connecting the compartments; (C) Immunofluorescence image of neurite structure following the microchannel placement.
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Figure 3: Structural connectivity between the two compartmentalized neuronal sub-populations. (A) Phase contrast image of neurite ladder structure intact after the removal of PDMS structures from the MEA surface; (B) Transfection image of a neurite grown across the microchannels connecting the compartments; (C) Immunofluorescence image of neurite structure following the microchannel placement.

Mentions: Visual observation of the culture over the developmental period indicates neurite growth across the compartment from DIV 3 onward. Neurites were observed to cross-over to the adjacent compartment through the microchannels along the full length of the compartment. Phase contrast imaging of cell bodies isolated within a compartment and neurites crossing-over to the adjacent compartment confirmed the physical confinement obtained by means of the microchannel structure (Figure 3).


Dual-compartment neurofluidic system for electrophysiological measurements in physically segregated and functionally connected neuronal cell culture.

Kanagasabapathi TT, Ciliberti D, Martinoia S, Wadman WJ, Decré MM - Front Neuroeng (2011)

Structural connectivity between the two compartmentalized neuronal sub-populations. (A) Phase contrast image of neurite ladder structure intact after the removal of PDMS structures from the MEA surface; (B) Transfection image of a neurite grown across the microchannels connecting the compartments; (C) Immunofluorescence image of neurite structure following the microchannel placement.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198030&req=5

Figure 3: Structural connectivity between the two compartmentalized neuronal sub-populations. (A) Phase contrast image of neurite ladder structure intact after the removal of PDMS structures from the MEA surface; (B) Transfection image of a neurite grown across the microchannels connecting the compartments; (C) Immunofluorescence image of neurite structure following the microchannel placement.
Mentions: Visual observation of the culture over the developmental period indicates neurite growth across the compartment from DIV 3 onward. Neurites were observed to cross-over to the adjacent compartment through the microchannels along the full length of the compartment. Phase contrast imaging of cell bodies isolated within a compartment and neurites crossing-over to the adjacent compartment confirmed the physical confinement obtained by means of the microchannel structure (Figure 3).

Bottom Line: Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated.The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining.In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques.

View Article: PubMed Central - PubMed

Affiliation: Minimally Invasive Healthcare Department, Philips Research Laboratories Eindhoven, Netherlands.

ABSTRACT
We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels.

No MeSH data available.


Related in: MedlinePlus