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

Shigeki Furuya, Asami Makino, Yoshio Hirabayashi

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)192-198
Number of pages7
JournalBrain Research Protocols
Volume3
Issue number2
DOIs
Publication statusPublished - Nov 1 1998

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Purkinje Cells
Dendrites
Eagles
Dendritic Cells
Serum-Free Culture Media
Nerve Growth Factors
Cerebellum
Culture Media
Suspensions
Cell Culture Techniques
Maintenance
Ions
Calcium
Growth
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

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An improved method for culturing cerebellar Purkinje cells with differentiated dendrites under a mixed monolayer setting. / Furuya, Shigeki; Makino, Asami; Hirabayashi, Yoshio.

In: Brain Research Protocols, Vol. 3, No. 2, 01.11.1998, p. 192-198.

Research output: Contribution to journalArticle

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