An improved method for culturing cerebellar Purkinje cells with differentiated dendrites under a mixed monolayer setting

Shigeki Furuya, Asami Makino, Yoshio Hirabayashi

研究成果: ジャーナルへの寄稿学術誌査読

101 被引用数 (Scopus)

抄録

We report here a novel cell culture protocol which facilitates in vitro survival and dendritic differentiation of cerebellar Purkinje cells in a monolayer, mixed culture setting. We found that the type of culture medium is a critical factor for the maintenance of these cells. Purkinje cells present in the single cell suspension of embryonic rat cerebellum were best maintained in a medium based on Dulbecco's modified Eagle's medium (DMEM)/F- 12 without the addition of known neurotrophic factors. These cells maintained in DMEM/F-12-based media displayed an approximately 2.5-3.5-fold increase in survival compared with cells maintained in the widely used Basal Medium Eagle's (BME)-based serum-free culture medium with the same supplements. This novel protocol permits not only enhanced survival but also accelerated, improved dendritic differentiation of these cells. Purkinje cells developed highly branched spiny dendrites by 14-16 days in vitro, which matches the time course of the dendritic growth of these cells in vivo. The Purkinje cells expressed metabotropic glutamate receptor 1α in the cell bodies and branched dendrites, and the intradendritic calcium concentration increased when trans-ACPD, a selective agonist of this receptor, was applied. This novel protocol allows the development of functional branched dendrites and therefore is useful for electrophysiological and ion-imaging studies on dendrites of Purkinje cells grown in vitro.

本文言語英語
ページ(範囲)192-198
ページ数7
ジャーナルBrain Research Protocols
3
2
DOI
出版ステータス出版済み - 11月 1998
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 神経科学(全般)

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