Differentiation is induced in three-dimensional cultures of brain cells immortalized by the LAP mammalian regulatory system

M. Toth, T. Shenk, E. Madarász

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Immortalized neuroectodermal precursor cell lines were generated from mouse brain by the SV40 large T antigen expressed under the control of the LAP (lac activating protein) mammalian regulatory system. The LAP system permits the reversible expression of T antigen as a function of the exogenous inducer, isopropyl-β-D-thiogalactopyranoside. Immortalized cells can be stably maintained in an undifferentiated state in monolayer cultures. Cell lines expressed the early neurofilament-like protein nestin, but not markers characteristic for mature cells such as the neurofilament light protein and glial fibrillary acidic protein. Downregulating the LAP-controlled T antigen with isopropyl-β-D-thiogalactopyranoside was not sufficient to induce differentiation. However, when cells formed three-dimensional aggregates, differentiation to a neuronal phenotype occurred, indicating that cell-cell interaction plays an important role in their differentiation. Cells in aggregates did not proliferate, even in the presence of T antigen, suggesting that an aggregation-induced signal to cease growth was dominant over the growth signal of T antigen. Further morphological differentiation was induced by basic fibroblast growth factor. These immortalized cells should facilitate molecular and cellular studies concerned with the mechanism of commitment, fate determination, and mitotic arrest of neuronal precursor cells in the developing mammalian CNS.

Original languageEnglish
Pages (from-to)764-774
Number of pages11
JournalJournal of Neuroscience Research
Volume41
Issue number6
DOIs
Publication statusPublished - 1995

Fingerprint

Viral Tumor Antigens
Cell Culture Techniques
Brain
Thiogalactosides
Proteins
Polyomavirus Transforming Antigens
Neurofilament Proteins
Cell Line
Nestin
Glial Fibrillary Acidic Protein
Fibroblast Growth Factor 2
Growth
Cell Communication
Down-Regulation
Phenotype

Keywords

  • aggregation
  • nestin
  • neurofilament
  • repressor
  • T antigen

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Differentiation is induced in three-dimensional cultures of brain cells immortalized by the LAP mammalian regulatory system. / Toth, M.; Shenk, T.; Madarász, E.

In: Journal of Neuroscience Research, Vol. 41, No. 6, 1995, p. 764-774.

Research output: Contribution to journalArticle

@article{7a2da84dc82141b3b0bc13ded3413de0,
title = "Differentiation is induced in three-dimensional cultures of brain cells immortalized by the LAP mammalian regulatory system",
abstract = "Immortalized neuroectodermal precursor cell lines were generated from mouse brain by the SV40 large T antigen expressed under the control of the LAP (lac activating protein) mammalian regulatory system. The LAP system permits the reversible expression of T antigen as a function of the exogenous inducer, isopropyl-β-D-thiogalactopyranoside. Immortalized cells can be stably maintained in an undifferentiated state in monolayer cultures. Cell lines expressed the early neurofilament-like protein nestin, but not markers characteristic for mature cells such as the neurofilament light protein and glial fibrillary acidic protein. Downregulating the LAP-controlled T antigen with isopropyl-β-D-thiogalactopyranoside was not sufficient to induce differentiation. However, when cells formed three-dimensional aggregates, differentiation to a neuronal phenotype occurred, indicating that cell-cell interaction plays an important role in their differentiation. Cells in aggregates did not proliferate, even in the presence of T antigen, suggesting that an aggregation-induced signal to cease growth was dominant over the growth signal of T antigen. Further morphological differentiation was induced by basic fibroblast growth factor. These immortalized cells should facilitate molecular and cellular studies concerned with the mechanism of commitment, fate determination, and mitotic arrest of neuronal precursor cells in the developing mammalian CNS.",
keywords = "aggregation, nestin, neurofilament, repressor, T antigen",
author = "M. Toth and T. Shenk and E. Madar{\'a}sz",
year = "1995",
doi = "10.1002/jnr.490410607",
language = "English",
volume = "41",
pages = "764--774",
journal = "Journal of Neuroscience Research",
issn = "0360-4012",
publisher = "Wiley-Liss Inc.",
number = "6",

}

TY - JOUR

T1 - Differentiation is induced in three-dimensional cultures of brain cells immortalized by the LAP mammalian regulatory system

AU - Toth, M.

AU - Shenk, T.

AU - Madarász, E.

PY - 1995

Y1 - 1995

N2 - Immortalized neuroectodermal precursor cell lines were generated from mouse brain by the SV40 large T antigen expressed under the control of the LAP (lac activating protein) mammalian regulatory system. The LAP system permits the reversible expression of T antigen as a function of the exogenous inducer, isopropyl-β-D-thiogalactopyranoside. Immortalized cells can be stably maintained in an undifferentiated state in monolayer cultures. Cell lines expressed the early neurofilament-like protein nestin, but not markers characteristic for mature cells such as the neurofilament light protein and glial fibrillary acidic protein. Downregulating the LAP-controlled T antigen with isopropyl-β-D-thiogalactopyranoside was not sufficient to induce differentiation. However, when cells formed three-dimensional aggregates, differentiation to a neuronal phenotype occurred, indicating that cell-cell interaction plays an important role in their differentiation. Cells in aggregates did not proliferate, even in the presence of T antigen, suggesting that an aggregation-induced signal to cease growth was dominant over the growth signal of T antigen. Further morphological differentiation was induced by basic fibroblast growth factor. These immortalized cells should facilitate molecular and cellular studies concerned with the mechanism of commitment, fate determination, and mitotic arrest of neuronal precursor cells in the developing mammalian CNS.

AB - Immortalized neuroectodermal precursor cell lines were generated from mouse brain by the SV40 large T antigen expressed under the control of the LAP (lac activating protein) mammalian regulatory system. The LAP system permits the reversible expression of T antigen as a function of the exogenous inducer, isopropyl-β-D-thiogalactopyranoside. Immortalized cells can be stably maintained in an undifferentiated state in monolayer cultures. Cell lines expressed the early neurofilament-like protein nestin, but not markers characteristic for mature cells such as the neurofilament light protein and glial fibrillary acidic protein. Downregulating the LAP-controlled T antigen with isopropyl-β-D-thiogalactopyranoside was not sufficient to induce differentiation. However, when cells formed three-dimensional aggregates, differentiation to a neuronal phenotype occurred, indicating that cell-cell interaction plays an important role in their differentiation. Cells in aggregates did not proliferate, even in the presence of T antigen, suggesting that an aggregation-induced signal to cease growth was dominant over the growth signal of T antigen. Further morphological differentiation was induced by basic fibroblast growth factor. These immortalized cells should facilitate molecular and cellular studies concerned with the mechanism of commitment, fate determination, and mitotic arrest of neuronal precursor cells in the developing mammalian CNS.

KW - aggregation

KW - nestin

KW - neurofilament

KW - repressor

KW - T antigen

UR - http://www.scopus.com/inward/record.url?scp=0029122790&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029122790&partnerID=8YFLogxK

U2 - 10.1002/jnr.490410607

DO - 10.1002/jnr.490410607

M3 - Article

VL - 41

SP - 764

EP - 774

JO - Journal of Neuroscience Research

JF - Journal of Neuroscience Research

SN - 0360-4012

IS - 6

ER -