Optimization of oligodendrocyte progenitor cell culture method for enhanced survival

Zhongshu Yang, Masahiko Watanabe, Akiko Nishiyama

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Oligodendrocyte progenitor cells (OPCs, NG2 glia) play an important role not only as progenitor cells that give rise to myelinating cells in the central nervous system (CNS), but also as an active participant in the neural network. It is necessary to develop a simplified method for generating large quantities of highly purified OPCs for biochemical studies and to establish a neuron-OPC coculture method for functional studies on the mechanism of neuron-OPC signaling. In this study, we have compared the effects of plating density and culture medium on purity, survival, and differentiation of cells collected from primary rat mixed glial cultures by differential adhesion. Comparison of two chemically-defined culture media, Dulbecco's modified Eagle's medium with N1 supplements (N1/DMEM) and Neurobasal medium with B27 supplements (B27/NBM) revealed that while both media successfully maintained greater than 90% pure OPCs after 3 days, B27/NBM was significantly more effective in maintaining viable cells and in supporting oligodendrocyte differentiation than N1/DMEM, and this effect was more pronounced in low density cultures. Furthermore, B27/NBM supported neuron-OPC coculture in which OPCs remained as NG2-positive progenitors and neurons differentiated to form synapses over a period of 3 weeks.

Original languageEnglish (US)
Pages (from-to)50-56
Number of pages7
JournalJournal of Neuroscience Methods
Volume149
Issue number1
DOIs
StatePublished - Nov 30 2005
Externally publishedYes

Fingerprint

Oligodendroglia
Stem Cells
Cell Culture Techniques
Neurons
Coculture Techniques
Neuroglia
Culture Media
Eagles
Synapses
Cell Differentiation
Cell Survival
Central Nervous System

Keywords

  • Differentiation
  • Glia
  • Hippocampal neuron
  • N1 medium
  • Neurobasal medium
  • NG2
  • Oligodendrocyte progenitor

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Optimization of oligodendrocyte progenitor cell culture method for enhanced survival. / Yang, Zhongshu; Watanabe, Masahiko; Nishiyama, Akiko.

In: Journal of Neuroscience Methods, Vol. 149, No. 1, 30.11.2005, p. 50-56.

Research output: Contribution to journalArticle

Yang, Zhongshu ; Watanabe, Masahiko ; Nishiyama, Akiko. / Optimization of oligodendrocyte progenitor cell culture method for enhanced survival. In: Journal of Neuroscience Methods. 2005 ; Vol. 149, No. 1. pp. 50-56.
@article{95ad3c98d57a4b598f292898566d7d4c,
title = "Optimization of oligodendrocyte progenitor cell culture method for enhanced survival",
abstract = "Oligodendrocyte progenitor cells (OPCs, NG2 glia) play an important role not only as progenitor cells that give rise to myelinating cells in the central nervous system (CNS), but also as an active participant in the neural network. It is necessary to develop a simplified method for generating large quantities of highly purified OPCs for biochemical studies and to establish a neuron-OPC coculture method for functional studies on the mechanism of neuron-OPC signaling. In this study, we have compared the effects of plating density and culture medium on purity, survival, and differentiation of cells collected from primary rat mixed glial cultures by differential adhesion. Comparison of two chemically-defined culture media, Dulbecco's modified Eagle's medium with N1 supplements (N1/DMEM) and Neurobasal medium with B27 supplements (B27/NBM) revealed that while both media successfully maintained greater than 90{\%} pure OPCs after 3 days, B27/NBM was significantly more effective in maintaining viable cells and in supporting oligodendrocyte differentiation than N1/DMEM, and this effect was more pronounced in low density cultures. Furthermore, B27/NBM supported neuron-OPC coculture in which OPCs remained as NG2-positive progenitors and neurons differentiated to form synapses over a period of 3 weeks.",
keywords = "Differentiation, Glia, Hippocampal neuron, N1 medium, Neurobasal medium, NG2, Oligodendrocyte progenitor",
author = "Zhongshu Yang and Masahiko Watanabe and Akiko Nishiyama",
year = "2005",
month = "11",
day = "30",
doi = "10.1016/j.jneumeth.2005.05.003",
language = "English (US)",
volume = "149",
pages = "50--56",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Optimization of oligodendrocyte progenitor cell culture method for enhanced survival

AU - Yang, Zhongshu

AU - Watanabe, Masahiko

AU - Nishiyama, Akiko

PY - 2005/11/30

Y1 - 2005/11/30

N2 - Oligodendrocyte progenitor cells (OPCs, NG2 glia) play an important role not only as progenitor cells that give rise to myelinating cells in the central nervous system (CNS), but also as an active participant in the neural network. It is necessary to develop a simplified method for generating large quantities of highly purified OPCs for biochemical studies and to establish a neuron-OPC coculture method for functional studies on the mechanism of neuron-OPC signaling. In this study, we have compared the effects of plating density and culture medium on purity, survival, and differentiation of cells collected from primary rat mixed glial cultures by differential adhesion. Comparison of two chemically-defined culture media, Dulbecco's modified Eagle's medium with N1 supplements (N1/DMEM) and Neurobasal medium with B27 supplements (B27/NBM) revealed that while both media successfully maintained greater than 90% pure OPCs after 3 days, B27/NBM was significantly more effective in maintaining viable cells and in supporting oligodendrocyte differentiation than N1/DMEM, and this effect was more pronounced in low density cultures. Furthermore, B27/NBM supported neuron-OPC coculture in which OPCs remained as NG2-positive progenitors and neurons differentiated to form synapses over a period of 3 weeks.

AB - Oligodendrocyte progenitor cells (OPCs, NG2 glia) play an important role not only as progenitor cells that give rise to myelinating cells in the central nervous system (CNS), but also as an active participant in the neural network. It is necessary to develop a simplified method for generating large quantities of highly purified OPCs for biochemical studies and to establish a neuron-OPC coculture method for functional studies on the mechanism of neuron-OPC signaling. In this study, we have compared the effects of plating density and culture medium on purity, survival, and differentiation of cells collected from primary rat mixed glial cultures by differential adhesion. Comparison of two chemically-defined culture media, Dulbecco's modified Eagle's medium with N1 supplements (N1/DMEM) and Neurobasal medium with B27 supplements (B27/NBM) revealed that while both media successfully maintained greater than 90% pure OPCs after 3 days, B27/NBM was significantly more effective in maintaining viable cells and in supporting oligodendrocyte differentiation than N1/DMEM, and this effect was more pronounced in low density cultures. Furthermore, B27/NBM supported neuron-OPC coculture in which OPCs remained as NG2-positive progenitors and neurons differentiated to form synapses over a period of 3 weeks.

KW - Differentiation

KW - Glia

KW - Hippocampal neuron

KW - N1 medium

KW - Neurobasal medium

KW - NG2

KW - Oligodendrocyte progenitor

UR - http://www.scopus.com/inward/record.url?scp=27744569894&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27744569894&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2005.05.003

DO - 10.1016/j.jneumeth.2005.05.003

M3 - Article

C2 - 15975663

AN - SCOPUS:27744569894

VL - 149

SP - 50

EP - 56

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 1

ER -