Long-term survival of cortical neurones from adult guinea-pig maintained in low-density cultures

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In vitro survival of neurones isolated from adult mammalian brain is normally scarce and the postnatal age limit for obtaining viable cultures of cortical, hippocampal and diencephalic neurones is commonly two weeks. Here we describe a novel procedure for the establishment and long-term maintenance of cortical neurones of the adult mammalian brain in low-density cultures. Neurones isolated from the piriform cortex of 30- to 90-day-old guinea-pigs were initially grown in a chemically defined medium enriched with basic fibroblast growth factor (bFGF); later, a small quantity of foetal bovine serum (FBS) was added to facilitate cell differentiation. Under these conditions cells could be maintained in culture for at least 3 weeks, when indirect immunocytochemistry and whole-cell patch-clamp recordings were performed. Cells exhibiting neuronal morphology expressed the neuronal marker microtubule associated protein-2 (MAP2) and generated action potentials. Moreover, about 70% of the MAP2-immunoreactive cells were simultaneously labelled with anti-γ-aminobutyric acid (GABA) antibody. Cells expressing neuronal antigens were never labelled by antibody raised against the glial marker glial fibrillary acidic protein (GFAP). These results indicate that long-term survival of adult neurones can be achieved under definite culture conditions.

Original languageEnglish
Pages (from-to)1559-1564
Number of pages6
JournalNeuroReport
Volume7
Issue number10
Publication statusPublished - 1996

Fingerprint

Guinea Pigs
Neurons
Microtubule-Associated Proteins
Aminobutyrates
Antibodies
Glial Fibrillary Acidic Protein
Brain
Fibroblast Growth Factor 2
Neuroglia
gamma-Aminobutyric Acid
Action Potentials
Cell Differentiation
Immunohistochemistry
Antigens
Serum

Keywords

  • BFGF
  • Cultures from adult mammalian brain
  • Neuronal cultures
  • Piriform cortex

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

@article{5d3ed7d49a2742659b30fbbcdd4ddc77,
title = "Long-term survival of cortical neurones from adult guinea-pig maintained in low-density cultures",
abstract = "In vitro survival of neurones isolated from adult mammalian brain is normally scarce and the postnatal age limit for obtaining viable cultures of cortical, hippocampal and diencephalic neurones is commonly two weeks. Here we describe a novel procedure for the establishment and long-term maintenance of cortical neurones of the adult mammalian brain in low-density cultures. Neurones isolated from the piriform cortex of 30- to 90-day-old guinea-pigs were initially grown in a chemically defined medium enriched with basic fibroblast growth factor (bFGF); later, a small quantity of foetal bovine serum (FBS) was added to facilitate cell differentiation. Under these conditions cells could be maintained in culture for at least 3 weeks, when indirect immunocytochemistry and whole-cell patch-clamp recordings were performed. Cells exhibiting neuronal morphology expressed the neuronal marker microtubule associated protein-2 (MAP2) and generated action potentials. Moreover, about 70{\%} of the MAP2-immunoreactive cells were simultaneously labelled with anti-γ-aminobutyric acid (GABA) antibody. Cells expressing neuronal antigens were never labelled by antibody raised against the glial marker glial fibrillary acidic protein (GFAP). These results indicate that long-term survival of adult neurones can be achieved under definite culture conditions.",
keywords = "BFGF, Cultures from adult mammalian brain, Neuronal cultures, Piriform cortex",
author = "Jacopo Magistretti and {De Curtis}, Marco and Angelo Vescovi and Rossella Galli and Angela Gritti",
year = "1996",
language = "English",
volume = "7",
pages = "1559--1564",
journal = "NeuroReport",
issn = "0959-4965",
publisher = "Lippincott Williams and Wilkins",
number = "10",

}

TY - JOUR

T1 - Long-term survival of cortical neurones from adult guinea-pig maintained in low-density cultures

AU - Magistretti, Jacopo

AU - De Curtis, Marco

AU - Vescovi, Angelo

AU - Galli, Rossella

AU - Gritti, Angela

PY - 1996

Y1 - 1996

N2 - In vitro survival of neurones isolated from adult mammalian brain is normally scarce and the postnatal age limit for obtaining viable cultures of cortical, hippocampal and diencephalic neurones is commonly two weeks. Here we describe a novel procedure for the establishment and long-term maintenance of cortical neurones of the adult mammalian brain in low-density cultures. Neurones isolated from the piriform cortex of 30- to 90-day-old guinea-pigs were initially grown in a chemically defined medium enriched with basic fibroblast growth factor (bFGF); later, a small quantity of foetal bovine serum (FBS) was added to facilitate cell differentiation. Under these conditions cells could be maintained in culture for at least 3 weeks, when indirect immunocytochemistry and whole-cell patch-clamp recordings were performed. Cells exhibiting neuronal morphology expressed the neuronal marker microtubule associated protein-2 (MAP2) and generated action potentials. Moreover, about 70% of the MAP2-immunoreactive cells were simultaneously labelled with anti-γ-aminobutyric acid (GABA) antibody. Cells expressing neuronal antigens were never labelled by antibody raised against the glial marker glial fibrillary acidic protein (GFAP). These results indicate that long-term survival of adult neurones can be achieved under definite culture conditions.

AB - In vitro survival of neurones isolated from adult mammalian brain is normally scarce and the postnatal age limit for obtaining viable cultures of cortical, hippocampal and diencephalic neurones is commonly two weeks. Here we describe a novel procedure for the establishment and long-term maintenance of cortical neurones of the adult mammalian brain in low-density cultures. Neurones isolated from the piriform cortex of 30- to 90-day-old guinea-pigs were initially grown in a chemically defined medium enriched with basic fibroblast growth factor (bFGF); later, a small quantity of foetal bovine serum (FBS) was added to facilitate cell differentiation. Under these conditions cells could be maintained in culture for at least 3 weeks, when indirect immunocytochemistry and whole-cell patch-clamp recordings were performed. Cells exhibiting neuronal morphology expressed the neuronal marker microtubule associated protein-2 (MAP2) and generated action potentials. Moreover, about 70% of the MAP2-immunoreactive cells were simultaneously labelled with anti-γ-aminobutyric acid (GABA) antibody. Cells expressing neuronal antigens were never labelled by antibody raised against the glial marker glial fibrillary acidic protein (GFAP). These results indicate that long-term survival of adult neurones can be achieved under definite culture conditions.

KW - BFGF

KW - Cultures from adult mammalian brain

KW - Neuronal cultures

KW - Piriform cortex

UR - http://www.scopus.com/inward/record.url?scp=0029784362&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029784362&partnerID=8YFLogxK

M3 - Article

C2 - 8904755

AN - SCOPUS:0029784362

VL - 7

SP - 1559

EP - 1564

JO - NeuroReport

JF - NeuroReport

SN - 0959-4965

IS - 10

ER -