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Cellular and molecular neurobiology
Oct, 2022;42 (7): 2337-2353.

A Novel Methodology Using Dexamethasone to Induce Neuronal Differentiation in the CNS-Derived Catecholaminergic CAD Cells.

Ekkaphot Khongkla, Kwanchanok Uppakara, Nittaya Boonmuen, Kanit Bhukhai, Witchuda Saengsawang
Author Information
  1. Ekkaphot Khongkla: Department of Physiology, Faculty of Science, Mahidol University, 272 Rama 6 Rd. Payathai, Ratchathewi district, Bangkok, 10400, Thailand.
  2. Kwanchanok Uppakara: Toxicology Graduate Program, Faculty of Sciene, Mahidol University, Bangkok, Thailand.
  3. Nittaya Boonmuen: Department of Physiology, Faculty of Science, Mahidol University, 272 Rama 6 Rd. Payathai, Ratchathewi district, Bangkok, 10400, Thailand.
  4. Kanit Bhukhai: Department of Physiology, Faculty of Science, Mahidol University, 272 Rama 6 Rd. Payathai, Ratchathewi district, Bangkok, 10400, Thailand.
  5. Witchuda Saengsawang: Department of Physiology, Faculty of Science, Mahidol University, 272 Rama 6 Rd. Payathai, Ratchathewi district, Bangkok, 10400, Thailand. witchudasaeng@gmail.com. ORCID
PMID: 34059943 DOI: 10.1007/s10571-021-01109-z

Abstract

The Cath.a-differentiated (CAD) cell line is a central nervous system-derived catecholaminergic cell line originating from tyrosine hydroxylase (TH)-producing neurons located around the locus coeruleus area of the mouse brain. CAD cells have been used as an in vitro model for cellular and molecular studies due to their ability to differentiate under serum-free media conditions. However, the lack of serum-derived survival factors, limits the longevity for differentiated CAD cells to be maintained in healthy conditions; thereby, limiting their use in long-term culture studies. Here, we present a novel differentiation method that utilizes dexamethasone (Dex), a synthetic glucocorticoid receptor agonist. Specifically, we discovered that the addition of 100 µM of Dex into the 1% fetal bovine serum (FBS)-supplemented media effectively induced neuronal differentiation of CAD cells, as characterized by neurite formation and elongation. Dex-differentiated CAD cells exited the cell cycle, stopped proliferating, extended the neurites, and expressed neuronal markers. These effects were dependent on the glucocorticoid receptors (GR) as they were abolished by GR knockdown. Importantly, Dex-differentiated CAD cells showed longer survival duration than serum-free differentiated CAD cells. In addition, RNA-sequencing and qPCR data demonstrate that several genes involved in proliferation, neuronal differentiation, and survival pathways were differentially expressed in the Dex-differentiated cells. This is the first study to reveal Dex as a novel differentiation methodology used to generate postmitotic neuronal CAD cells, which may be utilized as an in vitro neuronal model for cellular and molecular neurobiology research.

Keywords

CAD cells Cath.a-differentiated cell Dexamethasone Differentiation Neuronal cells line Proliferation

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Grants

  1. NDFR20/2563/Mahidol University
  2. IRN58W0004/Thailand Research Fund
  3. CIF/Faculty of Science, Mahidol University

MeSH Term

Animals
Cell Differentiation
Central Nervous System
Dexamethasone
Mice
Neurites
Neurons
Receptors, Glucocorticoid

Chemicals

Receptors, Glucocorticoid
Dexamethasone
Journal Article
Article has an altmetric score of 1

Word Cloud

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