Characterization of a murine mixed neuron-glia model and cellular responses to regulatory T cell-derived factors

Marie Dittmer, Andrew Young, Thomas O'Hagan, George Eleftheriadis, Peter Bankhead, Yvonne Dombrowski, Reinhold J Medina, Denise C Fitzgerald

Research output: Contribution to journalArticle

2 Citations (Scopus)
223 Downloads (Pure)

Abstract

One of the unmet clinical needs in demyelinating diseases such as Multiple Sclerosis (MS) is to provide therapies that actively enhance the process of myelin regeneration (remyelination) in the central nervous system (CNS). Oligodendrocytes, the myelinating cells of the CNS, play a central role in remyelination and originate from oligodendrocyte progenitor cells (OPCs). We recently showed that depletion of regulatory T cells (Treg) impairs remyelination in vivo, and that Treg-secreted factors directly enhance oligodendrocyte differentiation. Here we aim to further characterize the dynamics of Treg-enhanced oligodendrocyte differentiation as well as elucidate the cellular components of a murine mixed neuron-glia model. Murine mixed neuron-glia cultures were generated from P2-7 C57BL/6 mice and characterized for percentage of neuronal and glial cell populations prior to treatment at 7 days in vitro (div) as well as after treatment with Treg-conditioned media at multiple timepoints up to 12 div. Mixed neuron-glia cultures consisted of approximately 30% oligodendroglial lineage cells, 20% neurons and 10% microglia. Furthermore, a full layer of astrocytes, that could not be quantified, was present. Treatment with Treg-conditioned media enhanced the proportion of MBP+ oligodendrocytes and decreased the proportion of PDGFRα+ OPCs, but did not affect OPC proliferation or survival. Treg-enhanced oligodendrocyte differentiation was not caused by Treg polarizing factors, was dependent on the number of activation cycles Treg underwent and was robustly achieved by using 5% conditioned media. These studies provide in-depth characterization of a murine mixed neuron-glia model as well as further insights into the dynamics of Treg-enhanced oligodendrocyte differentiation.

Original languageEnglish
Pages (from-to)1-25
JournalMolecular Brain
Volume11
Issue number1
DOIs
Publication statusPublished - 02 May 2018

Fingerprint

TCF Transcription Factors
Oligodendroglia
Regulatory T-Lymphocytes
Neuroglia
Neurons
Conditioned Culture Medium
Stem Cells
Central Nervous System
Microglia
Demyelinating Diseases
Myelin Sheath
Inbred C57BL Mouse
Astrocytes
Multiple Sclerosis
Regeneration
Cell Survival
Cell Proliferation

Keywords

  • Journal Article

Cite this

@article{6fb86d513b23427ab617b1c9fc6c217f,
title = "Characterization of a murine mixed neuron-glia model and cellular responses to regulatory T cell-derived factors",
abstract = "One of the unmet clinical needs in demyelinating diseases such as Multiple Sclerosis (MS) is to provide therapies that actively enhance the process of myelin regeneration (remyelination) in the central nervous system (CNS). Oligodendrocytes, the myelinating cells of the CNS, play a central role in remyelination and originate from oligodendrocyte progenitor cells (OPCs). We recently showed that depletion of regulatory T cells (Treg) impairs remyelination in vivo, and that Treg-secreted factors directly enhance oligodendrocyte differentiation. Here we aim to further characterize the dynamics of Treg-enhanced oligodendrocyte differentiation as well as elucidate the cellular components of a murine mixed neuron-glia model. Murine mixed neuron-glia cultures were generated from P2-7 C57BL/6 mice and characterized for percentage of neuronal and glial cell populations prior to treatment at 7 days in vitro (div) as well as after treatment with Treg-conditioned media at multiple timepoints up to 12 div. Mixed neuron-glia cultures consisted of approximately 30{\%} oligodendroglial lineage cells, 20{\%} neurons and 10{\%} microglia. Furthermore, a full layer of astrocytes, that could not be quantified, was present. Treatment with Treg-conditioned media enhanced the proportion of MBP+ oligodendrocytes and decreased the proportion of PDGFRα+ OPCs, but did not affect OPC proliferation or survival. Treg-enhanced oligodendrocyte differentiation was not caused by Treg polarizing factors, was dependent on the number of activation cycles Treg underwent and was robustly achieved by using 5{\%} conditioned media. These studies provide in-depth characterization of a murine mixed neuron-glia model as well as further insights into the dynamics of Treg-enhanced oligodendrocyte differentiation.",
keywords = "Journal Article",
author = "Marie Dittmer and Andrew Young and Thomas O'Hagan and George Eleftheriadis and Peter Bankhead and Yvonne Dombrowski and Medina, {Reinhold J} and Fitzgerald, {Denise C}",
year = "2018",
month = "5",
day = "2",
doi = "10.1186/s13041-018-0367-6",
language = "English",
volume = "11",
pages = "1--25",
journal = "Molecular Brain",
issn = "1756-6606",
publisher = "BioMed Central",
number = "1",

}

Characterization of a murine mixed neuron-glia model and cellular responses to regulatory T cell-derived factors. / Dittmer, Marie; Young, Andrew; O'Hagan, Thomas; Eleftheriadis, George; Bankhead, Peter; Dombrowski, Yvonne; Medina, Reinhold J; Fitzgerald, Denise C.

In: Molecular Brain, Vol. 11, No. 1, 02.05.2018, p. 1-25.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of a murine mixed neuron-glia model and cellular responses to regulatory T cell-derived factors

AU - Dittmer, Marie

AU - Young, Andrew

AU - O'Hagan, Thomas

AU - Eleftheriadis, George

AU - Bankhead, Peter

AU - Dombrowski, Yvonne

AU - Medina, Reinhold J

AU - Fitzgerald, Denise C

PY - 2018/5/2

Y1 - 2018/5/2

N2 - One of the unmet clinical needs in demyelinating diseases such as Multiple Sclerosis (MS) is to provide therapies that actively enhance the process of myelin regeneration (remyelination) in the central nervous system (CNS). Oligodendrocytes, the myelinating cells of the CNS, play a central role in remyelination and originate from oligodendrocyte progenitor cells (OPCs). We recently showed that depletion of regulatory T cells (Treg) impairs remyelination in vivo, and that Treg-secreted factors directly enhance oligodendrocyte differentiation. Here we aim to further characterize the dynamics of Treg-enhanced oligodendrocyte differentiation as well as elucidate the cellular components of a murine mixed neuron-glia model. Murine mixed neuron-glia cultures were generated from P2-7 C57BL/6 mice and characterized for percentage of neuronal and glial cell populations prior to treatment at 7 days in vitro (div) as well as after treatment with Treg-conditioned media at multiple timepoints up to 12 div. Mixed neuron-glia cultures consisted of approximately 30% oligodendroglial lineage cells, 20% neurons and 10% microglia. Furthermore, a full layer of astrocytes, that could not be quantified, was present. Treatment with Treg-conditioned media enhanced the proportion of MBP+ oligodendrocytes and decreased the proportion of PDGFRα+ OPCs, but did not affect OPC proliferation or survival. Treg-enhanced oligodendrocyte differentiation was not caused by Treg polarizing factors, was dependent on the number of activation cycles Treg underwent and was robustly achieved by using 5% conditioned media. These studies provide in-depth characterization of a murine mixed neuron-glia model as well as further insights into the dynamics of Treg-enhanced oligodendrocyte differentiation.

AB - One of the unmet clinical needs in demyelinating diseases such as Multiple Sclerosis (MS) is to provide therapies that actively enhance the process of myelin regeneration (remyelination) in the central nervous system (CNS). Oligodendrocytes, the myelinating cells of the CNS, play a central role in remyelination and originate from oligodendrocyte progenitor cells (OPCs). We recently showed that depletion of regulatory T cells (Treg) impairs remyelination in vivo, and that Treg-secreted factors directly enhance oligodendrocyte differentiation. Here we aim to further characterize the dynamics of Treg-enhanced oligodendrocyte differentiation as well as elucidate the cellular components of a murine mixed neuron-glia model. Murine mixed neuron-glia cultures were generated from P2-7 C57BL/6 mice and characterized for percentage of neuronal and glial cell populations prior to treatment at 7 days in vitro (div) as well as after treatment with Treg-conditioned media at multiple timepoints up to 12 div. Mixed neuron-glia cultures consisted of approximately 30% oligodendroglial lineage cells, 20% neurons and 10% microglia. Furthermore, a full layer of astrocytes, that could not be quantified, was present. Treatment with Treg-conditioned media enhanced the proportion of MBP+ oligodendrocytes and decreased the proportion of PDGFRα+ OPCs, but did not affect OPC proliferation or survival. Treg-enhanced oligodendrocyte differentiation was not caused by Treg polarizing factors, was dependent on the number of activation cycles Treg underwent and was robustly achieved by using 5% conditioned media. These studies provide in-depth characterization of a murine mixed neuron-glia model as well as further insights into the dynamics of Treg-enhanced oligodendrocyte differentiation.

KW - Journal Article

U2 - 10.1186/s13041-018-0367-6

DO - 10.1186/s13041-018-0367-6

M3 - Article

C2 - 29720228

VL - 11

SP - 1

EP - 25

JO - Molecular Brain

JF - Molecular Brain

SN - 1756-6606

IS - 1

ER -