Rapid method for culturing embryonic neuron-glial cell cocultures

Åsa Fex Svenningsen, Wei-Song Shan, David R Colman, Liliana Pedraza

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric neurons is seen after 3 weeks (2 weeks in ascorbic acid), suggesting that basal lamina production is important even for glial ensheathment in the enteric nervous system. No overgrowth of fibroblasts or other nonneuronal cells was noted in any cultures, and myelination of the peripheral nervous system and central nervous system cultures was very robust.
OriginalsprogEngelsk
TidsskriftJournal of Neuroscience Research
Vol/bind72
Udgave nummer5
Sider (fra-til)565-73
Antal sider8
ISSN0360-4012
DOI
StatusUdgivet - 2003

Fingeraftryk

Coculture Techniques
Neuroglia
Neurons
Spinal Ganglia
Peripheral Nervous System
Embryonic Structures
Central Nervous System
Fibroblasts
Serum

Citer dette

Svenningsen, Åsa Fex ; Shan, Wei-Song ; Colman, David R ; Pedraza, Liliana. / Rapid method for culturing embryonic neuron-glial cell cocultures. I: Journal of Neuroscience Research. 2003 ; Bind 72, Nr. 5. s. 565-73.
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abstract = "A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric neurons is seen after 3 weeks (2 weeks in ascorbic acid), suggesting that basal lamina production is important even for glial ensheathment in the enteric nervous system. No overgrowth of fibroblasts or other nonneuronal cells was noted in any cultures, and myelination of the peripheral nervous system and central nervous system cultures was very robust.",
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Rapid method for culturing embryonic neuron-glial cell cocultures. / Svenningsen, Åsa Fex; Shan, Wei-Song; Colman, David R; Pedraza, Liliana.

I: Journal of Neuroscience Research, Bind 72, Nr. 5, 2003, s. 565-73.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Rapid method for culturing embryonic neuron-glial cell cocultures

AU - Svenningsen, Åsa Fex

AU - Shan, Wei-Song

AU - Colman, David R

AU - Pedraza, Liliana

N1 - Copyright 2003 Wiley-Liss, Inc.

PY - 2003

Y1 - 2003

N2 - A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric neurons is seen after 3 weeks (2 weeks in ascorbic acid), suggesting that basal lamina production is important even for glial ensheathment in the enteric nervous system. No overgrowth of fibroblasts or other nonneuronal cells was noted in any cultures, and myelination of the peripheral nervous system and central nervous system cultures was very robust.

AB - A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric neurons is seen after 3 weeks (2 weeks in ascorbic acid), suggesting that basal lamina production is important even for glial ensheathment in the enteric nervous system. No overgrowth of fibroblasts or other nonneuronal cells was noted in any cultures, and myelination of the peripheral nervous system and central nervous system cultures was very robust.

KW - Animals

KW - Axons

KW - Calcium-Binding Protein, Vitamin D-Dependent

KW - Cell Differentiation

KW - Cells, Cultured

KW - Cerebellum

KW - Coculture Techniques

KW - Culture Media, Serum-Free

KW - Enteric Nervous System

KW - Female

KW - Ganglia, Spinal

KW - Intermediate Filament Proteins

KW - Membrane Glycoproteins

KW - Myelin Basic Proteins

KW - Myelin Sheath

KW - Myelin-Associated Glycoprotein

KW - Nerve Tissue Proteins

KW - Neuroglia

KW - Neurons

KW - Oligodendroglia

KW - Rats

KW - Rats, Sprague-Dawley

KW - Schwann Cells

KW - Stem Cells

KW - Trypsin

U2 - 10.1002/jnr.10610

DO - 10.1002/jnr.10610

M3 - Journal article

VL - 72

SP - 565

EP - 573

JO - Journal of Neuroscience Research

JF - Journal of Neuroscience Research

SN - 0360-4012

IS - 5

ER -