Limits...
Networked neural spheroid by neuro-bundle mimicking nervous system created by topology effect.

Jeong GS, Chang JY, Park JS, Lee SA, Park D, Woo J, An H, Lee CJ, Lee SH - Mol Brain (2015)

Bottom Line: During neural-network formation, neural progenitor cells successfully differentiated into glial and neuronal cells.These cells secreted laminin, forming an extracellular matrix around the host and satellite spheroids.Electrical stimuli were transmitted between networked neurospheroids in the resulting nerve-like neural bundle, as detected by imaging Ca(2+) signals in responding cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, College of Health Science, Korea University, Seoul, 136-100, South Korea. arysu94@gmail.com.

ABSTRACT
In most animals, the nervous system consists of the central nervous system (CNS) and the peripheral nervous system (PNS), the latter of which connects the CNS to all parts of the body. Damage and/or malfunction of the nervous system causes serious pathologies, including neurodegenerative disorders, spinal cord injury, and Alzheimer's disease. Thus, not surprising, considerable research effort, both in vivo and in vitro, has been devoted to studying the nervous system and signal transmission through it. However, conventional in vitro cell culture systems do not enable control over diverse aspects of the neural microenvironment. Moreover, formation of certain nervous system growth patterns in vitro remains a challenge. In this study, we developed a deep hemispherical, microchannel-networked, concave array system and applied it to generate three-dimensional nerve-like neural bundles. The deep hemicylindrical channel network was easily fabricated by exploiting the meniscus induced by the surface tension of a liquid poly(dimethylsiloxane) (PDMS) prepolymer. Neurospheroids spontaneously aggregated in each deep concave microwell and were networked to neighboring spheroids through the deep hemicylindrical channel. Notably, two types of satellite spheroids also formed in deep hemispherical microchannels through self-aggregation and acted as an anchoring point to enhance formation of nerve-like networks with neighboring spheroids. During neural-network formation, neural progenitor cells successfully differentiated into glial and neuronal cells. These cells secreted laminin, forming an extracellular matrix around the host and satellite spheroids. Electrical stimuli were transmitted between networked neurospheroids in the resulting nerve-like neural bundle, as detected by imaging Ca(2+) signals in responding cells.

Show MeSH

Related in: MedlinePlus

Neural network formation in a deep HCWN system. (a) Concave channel networks were fabricated by exploiting the surface tension of a PDMS prepolymer, as follows: i) Prepolymer PDMS was poured onto a base mold (PDMS). ii) The prepolymer was removed from the base mold by wiping out liquid PDMS, forming a meniscus. iii) For deep HCWN plates, PDMS prepolymer left behind after step ii is removed by suctioning. iv) After curing, the base mold was used for fabrication of a prepolymer PDMS concave channel network. (b,c) SEM images of a cross-section of the concave channel network, showing connections of concave well arrays with concave channels in shallow HCWN plates (b) and deep HCWN plates (c). (d) Schematic view showing the dimensions of shallow and deep HCWN plates. (e) Schematic depiction of formation of a neural network, including satellite spheroids and neurite bundles, in a concave channel network.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4379946&req=5

Fig1: Neural network formation in a deep HCWN system. (a) Concave channel networks were fabricated by exploiting the surface tension of a PDMS prepolymer, as follows: i) Prepolymer PDMS was poured onto a base mold (PDMS). ii) The prepolymer was removed from the base mold by wiping out liquid PDMS, forming a meniscus. iii) For deep HCWN plates, PDMS prepolymer left behind after step ii is removed by suctioning. iv) After curing, the base mold was used for fabrication of a prepolymer PDMS concave channel network. (b,c) SEM images of a cross-section of the concave channel network, showing connections of concave well arrays with concave channels in shallow HCWN plates (b) and deep HCWN plates (c). (d) Schematic view showing the dimensions of shallow and deep HCWN plates. (e) Schematic depiction of formation of a neural network, including satellite spheroids and neurite bundles, in a concave channel network.

Mentions: Using a sweeping and suctioning process, we successfully fabricated shallow and deep hemicylindrical channels, and concave microwells from the PDMS base mold without the use of complicated devices or processes. For fabrication of shallow HCWN plates, only a sweeping process was employed, resulting in the generation of a shallow (~70 μm deep) hemicylindrical channel. The residual PDMS polymer was approximately 41.97 ± 3.57 mg (after sweeping) and 33.91 ± 2.75 mg (after suction) in the base mold (Additional file 1: Figure S1e). Figure 1b shows a side view of an SEM image of the shallow hemicylindrical channel and concave wells integrated in the HCWN system. Deep HCWN plates were prepared similarly, but with addition of a suctioning step to remove remnant PDMS polymer. After suctioning, the height of the hemicylindrical channel was almost the same as the rectangular channel height (~300 μm) of the PDMS base mold (Figure 1c). Figure 1d shows an illustration of the shape and dimensions of shallow and deep HCWN plates. As shown in the figures, the deeper hemicylindrical channel is suitable for generating satellite spheroids owing to the comparatively smooth joint of the deep channel (Figure 1e).Figure 1


Networked neural spheroid by neuro-bundle mimicking nervous system created by topology effect.

Jeong GS, Chang JY, Park JS, Lee SA, Park D, Woo J, An H, Lee CJ, Lee SH - Mol Brain (2015)

Neural network formation in a deep HCWN system. (a) Concave channel networks were fabricated by exploiting the surface tension of a PDMS prepolymer, as follows: i) Prepolymer PDMS was poured onto a base mold (PDMS). ii) The prepolymer was removed from the base mold by wiping out liquid PDMS, forming a meniscus. iii) For deep HCWN plates, PDMS prepolymer left behind after step ii is removed by suctioning. iv) After curing, the base mold was used for fabrication of a prepolymer PDMS concave channel network. (b,c) SEM images of a cross-section of the concave channel network, showing connections of concave well arrays with concave channels in shallow HCWN plates (b) and deep HCWN plates (c). (d) Schematic view showing the dimensions of shallow and deep HCWN plates. (e) Schematic depiction of formation of a neural network, including satellite spheroids and neurite bundles, in a concave channel network.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4379946&req=5

Fig1: Neural network formation in a deep HCWN system. (a) Concave channel networks were fabricated by exploiting the surface tension of a PDMS prepolymer, as follows: i) Prepolymer PDMS was poured onto a base mold (PDMS). ii) The prepolymer was removed from the base mold by wiping out liquid PDMS, forming a meniscus. iii) For deep HCWN plates, PDMS prepolymer left behind after step ii is removed by suctioning. iv) After curing, the base mold was used for fabrication of a prepolymer PDMS concave channel network. (b,c) SEM images of a cross-section of the concave channel network, showing connections of concave well arrays with concave channels in shallow HCWN plates (b) and deep HCWN plates (c). (d) Schematic view showing the dimensions of shallow and deep HCWN plates. (e) Schematic depiction of formation of a neural network, including satellite spheroids and neurite bundles, in a concave channel network.
Mentions: Using a sweeping and suctioning process, we successfully fabricated shallow and deep hemicylindrical channels, and concave microwells from the PDMS base mold without the use of complicated devices or processes. For fabrication of shallow HCWN plates, only a sweeping process was employed, resulting in the generation of a shallow (~70 μm deep) hemicylindrical channel. The residual PDMS polymer was approximately 41.97 ± 3.57 mg (after sweeping) and 33.91 ± 2.75 mg (after suction) in the base mold (Additional file 1: Figure S1e). Figure 1b shows a side view of an SEM image of the shallow hemicylindrical channel and concave wells integrated in the HCWN system. Deep HCWN plates were prepared similarly, but with addition of a suctioning step to remove remnant PDMS polymer. After suctioning, the height of the hemicylindrical channel was almost the same as the rectangular channel height (~300 μm) of the PDMS base mold (Figure 1c). Figure 1d shows an illustration of the shape and dimensions of shallow and deep HCWN plates. As shown in the figures, the deeper hemicylindrical channel is suitable for generating satellite spheroids owing to the comparatively smooth joint of the deep channel (Figure 1e).Figure 1

Bottom Line: During neural-network formation, neural progenitor cells successfully differentiated into glial and neuronal cells.These cells secreted laminin, forming an extracellular matrix around the host and satellite spheroids.Electrical stimuli were transmitted between networked neurospheroids in the resulting nerve-like neural bundle, as detected by imaging Ca(2+) signals in responding cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, College of Health Science, Korea University, Seoul, 136-100, South Korea. arysu94@gmail.com.

ABSTRACT
In most animals, the nervous system consists of the central nervous system (CNS) and the peripheral nervous system (PNS), the latter of which connects the CNS to all parts of the body. Damage and/or malfunction of the nervous system causes serious pathologies, including neurodegenerative disorders, spinal cord injury, and Alzheimer's disease. Thus, not surprising, considerable research effort, both in vivo and in vitro, has been devoted to studying the nervous system and signal transmission through it. However, conventional in vitro cell culture systems do not enable control over diverse aspects of the neural microenvironment. Moreover, formation of certain nervous system growth patterns in vitro remains a challenge. In this study, we developed a deep hemispherical, microchannel-networked, concave array system and applied it to generate three-dimensional nerve-like neural bundles. The deep hemicylindrical channel network was easily fabricated by exploiting the meniscus induced by the surface tension of a liquid poly(dimethylsiloxane) (PDMS) prepolymer. Neurospheroids spontaneously aggregated in each deep concave microwell and were networked to neighboring spheroids through the deep hemicylindrical channel. Notably, two types of satellite spheroids also formed in deep hemispherical microchannels through self-aggregation and acted as an anchoring point to enhance formation of nerve-like networks with neighboring spheroids. During neural-network formation, neural progenitor cells successfully differentiated into glial and neuronal cells. These cells secreted laminin, forming an extracellular matrix around the host and satellite spheroids. Electrical stimuli were transmitted between networked neurospheroids in the resulting nerve-like neural bundle, as detected by imaging Ca(2+) signals in responding cells.

Show MeSH
Related in: MedlinePlus