Anisotropically organized three-dimensional culture platform for reconstruction of a hippocampal neural network

Kim, So Hyun; Im, Sun Kyoung; Oh, Soo Jin; Jeong, Soon-Wook; Yoon, En Sup; Lee, C. Justin; Choi, Nakwon; Hur, Eun Mi

doi:10.1038/ncomms14346

Cited by 101 publications

(104 citation statements)

References 72 publications

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“…Whole-cell patch-clamp recordings were achieved by applying collagenase as previously described [5]. For tonic GABA measurement, whole-cell recordings were made from neurons.…”

Section: Methodsmentioning

confidence: 99%

Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture

Woo

Chun

et al. 2017

Exp Neurobiol

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Brain is a rich environment where neurons and glia interact with neighboring cells as well as extracellular matrix in three-dimensional (3D) space. Astrocytes, which are the most abundant cells in the mammalian brain, reside in 3D space and extend highly branched processes that form microdomains and contact synapses. It has been suggested that astrocytes cultured in 3D might be maintained in a less reactive state as compared to those growing in a traditional, two-dimensional (2D) monolayer culture. However, the functional characterization of the astrocytes in 3D culture has been lacking. Here we cocultured neurons and astrocytes in 3D and examined the morphological, molecular biological, and electrophysiological properties of the 3D-cultured hippocampal astrocytes. In our 3D neuron-astrocyte coculture, astrocytes showed a typical morphology of a small soma with many branches and exhibited a unique membrane property of passive conductance, more closely resembling their native in vivo counterparts. Moreover, we also induced reactive astrocytosis in culture by infecting with high-titer adenovirus to mimic pathophysiological conditions in vivo. Adenoviral infection induced morphological changes in astrocytes, increased passive conductance, and increased GABA content as well as tonic GABA release, which are characteristics of reactive gliosis. Together, our study presents a powerful in vitro model resembling both physiological and pathophysiological conditions in vivo, and thereby provides a versatile experimental tool for studying various neurological diseases that accompany reactive astrocytes.

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“…Whole-cell patch-clamp recordings were achieved by applying collagenase as previously described [5]. For tonic GABA measurement, whole-cell recordings were made from neurons.…”

Section: Methodsmentioning

confidence: 99%

Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture

Woo

Chun

et al. 2017

Exp Neurobiol

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“…[76][77][78][79] The thus induced temporal anisotropy can be fixed by in situ polymerization to afford an anisotropic hydrogel. [80][81][82][83] In some cases,the compression-or stretching-induced deformation remains in place without in situ polymerization owing to the plastic nature of the system. [84][85][86][87] As al imitation of this method, the precursor isotropic hydrogels must withstand large forces so that their gel networks undergo sufficient deformation.…”

Section: Hydrogels With Oriented Polymer-chain Networkmentioning

confidence: 99%

Synthesis of Anisotropic Hydrogels and Their Applications

2018

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Owing to their water-rich structures, which are similar to those of biological tissues, hydrogels have long been regarded as promising scaffolds for artificial tissues and organs. However, in terms of the structural anisotropy, most synthetic hydrogels are substantially different from biological systems. Synthetic hydrogels are usually composed of randomly oriented three-dimensional polymer networks whereas biological systems adopt anisotropic structures with hierarchically integrated building units. Such anisotropic structures often play essential roles in biological systems to exhibit particular functions. In this context, anisotropic hydrogels provide an entry point for exploring biomimetic applications of hydrogels. Reflecting these aspects, an increasing number of studies on anisotropic hydrogels have been reported recently. This Minireview highlights the use and perspectives of these anisotropic hydrogels, particularly focusing on their preparation, structures, and applications.

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“…Immunostaining was performed as described elsewhere [32,44]. Cells were fixed with pre-warmed 4% (w/v) paraformaldehyde (PFA), 0.15% glutaraldehyde, and 0.1% (w/v) Triton X-100 dissolved in phosphate buffered saline (PBS) for 15 min at 37 • C. After washing with PBS twice, cells were treated with 0.1% (w/v) sodium borohydride (Honeywell Fluka, 71321, Charlotte, North Carolina, United States) dissolved in PBS for 20 min at RT.…”

Section: Immunofluorescencementioning

confidence: 99%

A Role of Microtubules in Oligodendrocyte Differentiation

Lee

Hur

2020

IJMS

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Oligodendrocytes are specialized cells that myelinate axons in the central nervous system. Defects in oligodendrocyte function and failure to form or maintain myelin sheaths can cause a number of neurological disorders. Oligodendrocytes are differentiated from oligodendrocyte progenitor cells (OPCs), which extend several processes that contact, elaborate, and eventually wrap axonal segments to form multilayered myelin sheaths. These processes require extensive changes in the cytoarchitecture and must be regulated by reorganization of the cytoskeleton. Here, we established a simple protocol to isolate and differentiate mouse OPCs, and by using this method, we investigated a role of microtubules (MTs) in oligodendrocyte differentiation. Oligodendrocytes developed a complex network of MTs during differentiation, and treatment of differentiating oligodendrocytes with nanomolar concentrations of MT-targeting agents (MTAs) markedly affected oligodendrocyte survival and differentiation. We found that acute exposure to vincristine and nocodazole at early stages of oligodendrocyte differentiation markedly increased MT arborization and enhanced differentiation, whereas taxol and epothilone B treatment produced opposing outcomes. Furthermore, treatment of myelinating co-cultures of oligodendrocytes and neurons with nanomolar concentrations of MTAs at late stages of oligodendrocyte differentiation induced dysmyelination. Together, these results suggest that MTs play an important role in the survival, differentiation, and myelination of oligodendrocytes.

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Anisotropically organized three-dimensional culture platform for reconstruction of a hippocampal neural network

Cited by 101 publications

References 72 publications

Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture

Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture

Synthesis of Anisotropic Hydrogels and Their Applications

A Role of Microtubules in Oligodendrocyte Differentiation

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