Co-culture platform for neuron-astrocyte interaction using optogenetic modulation

Seoyoung Hwang, Yena Lee, Sang Beom Jun

Research output: Contribution to journalArticlepeer-review

Abstract

For decades, the role of glial cells has attracted attention in the neuroscience field. Particularly, although the astrocyte is the most abundant glial cell type, it was believed to function as a passive support cell. However, recent evidence suggests that astrocytes actively release various gliotransmitters and signaling entities that regulate the excitability of pre-and post-synaptic neurons in the brain. In this study, we optimized the ratio of astrocytes and neurons to investigate the interaction between astrocytes and neurons. To this end, postnatal day 0–1 rodent hippocampi were dissociated and cultured. The neuron–astrocyte ratio was monitored for up to 3 weeks after treating the cultures with 0, 1, and 5 µM of cytosine arabinoside (Ara-C) at DIV 2. Subsequently, from postnatal transgenic (TG) mouse expressing ChR2 on astrocytes, hippocampi were cultured on the microelectrode array (MEA) with the desired neuron–astrocyte ratio. The astrocyte was irradiated using a 473 nm blue laser for 30 s in a cycle of 10 Hz and electrophysiological recording was performed to verify the activities of neurons induced by the stimulated astrocytes. Astrocytes and neurons in both co-cultures increased at an identical ratio when treated with 1 µM Ara-C, whereas they decreased significantly when treated with 5 µM Ara-C. Particularly, the laser-stimulated astrocytes induced an increase in the frequency of neuronal activities and lasted after illumination. The proposed co-culture platform is expected to be used in experiments to investigate the network between astrocytes and neurons in vitro.

Original languageEnglish
Pages (from-to)401-411
Number of pages11
JournalBiomedical Engineering Letters
Volume12
Issue number4
DOIs
StatePublished - Nov 2022

Bibliographical note

Funding Information:
We thank Prof. Sung Joong Lee and Dr. Woo-Hyun Cho from the School of Dentistry, Seoul National University for offering transgenic mice and helpful comments during the writing of the manuscript. This research was supported by Convergent Technology R&D Program for Human Augmentation through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (NRF-2019M3C1B8090805) and by the Korea Medical Device Development Fund Grant funded by the Korea government (the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, the Ministry of Health & Welfare, the Ministry of Food and Drug Safety) (1711139110, KMDF_PR_20210527_0006).

Funding Information:
We thank Prof. Sung Joong Lee and Dr. Woo-Hyun Cho from the School of Dentistry, Seoul National University for offering transgenic mice and helpful comments during the writing of the manuscript. This research was supported by Convergent Technology R&D Program for Human Augmentation through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (NRF-2019M3C1B8090805) and by the Korea Medical Device Development Fund Grant funded by the Korea government (the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, the Ministry of Health & Welfare, the Ministry of Food and Drug Safety) (1711139110, KMDF_PR_20210527_0006).

Publisher Copyright:
© 2022, Korean Society of Medical and Biological Engineering.

Keywords

  • Astrocyte
  • Co-culture
  • Microelectrode array
  • Optogenetic modulation

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