View in EDS

Influence of rat substrain and growth conditions on the characteristics of primary cultures of adult rat spinal cord astrocytes

Saved in:
Bibliographic Details
Title: Influence of rat substrain and growth conditions on the characteristics of primary cultures of adult rat spinal cord astrocytes
Authors: Codeluppi, Simone1 simone.codeluppi@ki.se, Gregory, Ebba Norsted2 ebba.gregory@ki.se, Kjell, Jacob1 jacob.kjell@ki.se, Wigerblad, Gustaf2 gustaf.wigerblad@ki.se, Olson, Lars1 lars.olson@ki.se, Svensson, Camilla I.2 Camilla.svensson@ki.se
Source: Journal of Neuroscience Methods. Apr2011, Vol. 197 Issue 1, p118-127. 10p.
Subjects: Neural development, Cell culture, Astrocytes, Cellular signal transduction, Laboratory rats, Gene expression, Spinal cord injuries, Glutamine synthetase
Abstract: Abstract: Primary astrocyte cell cultures have become a valuable tool for studies of signaling pathways that regulate astrocyte physiology, reactivity, and function; however, differences in culture preparation affect data reproducibility. The aim of this work was to define optimal conditions for obtaining primary astrocytes from adult rat spinal cord with an expression profile most similar to adult human spinal cord astrocytes. Hence, we examined whether different Sprague–Dawley substrains and culture conditions affect astrocyte culture quality. Medium supplemented with fetal bovine serum from three sources (Sigma, Gibco, Hyclone) or a medium with defined composition (AM medium) was used to culture astrocytes isolated from spinal cords of adult Harlan and Charles River Spraque–Dawley rats. Purity was significantly different between cultures established in media with different sera. No microglia were detected in AM or Hyclone cultures. Gene expression was also affected, with AM cultures expressing the highest level of glutamine synthetase, connexin-43, and glutamate transporter-1. Interestingly, cell response to starvation was substrain dependent. Charles River-derived cultures responded the least, while astrocytes derived from Harlan rats showed a greater decrease in Gfap and glutamine synthetase, suggesting a more quiescent phenotype. Human and Harlan astrocytes cultured in AM media responded similarly to starvation. Taken together, this study shows that rat substrain and growth medium composition affect purity, expression profile and response to starvation of primary astrocytes suggesting that cultures of Harlan rats in AM media have optimal astrocyte characteristics, purity, and similarity to human astrocytes. [Copyright &y& Elsevier]
Copyright of Journal of Neuroscience Methods is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
FullText Text:
  Availability: 0
Header DbId: egs
DbLabel: Engineering Source
An: 59772851
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
IllustrationInfo
Items – Name: Title
  Label: Title
  Group: Ti
  Data: Influence of rat substrain and growth conditions on the characteristics of primary cultures of adult rat spinal cord astrocytes
– Name: Author
  Label: Authors
  Group: Au
  Data: <searchLink fieldCode="AR" term="%22Codeluppi%2C+Simone%22">Codeluppi, Simone</searchLink><relatesTo>1</relatesTo><i> simone.codeluppi@ki.se</i><br /><searchLink fieldCode="AR" term="%22Gregory%2C+Ebba+Norsted%22">Gregory, Ebba Norsted</searchLink><relatesTo>2</relatesTo><i> ebba.gregory@ki.se</i><br /><searchLink fieldCode="AR" term="%22Kjell%2C+Jacob%22">Kjell, Jacob</searchLink><relatesTo>1</relatesTo><i> jacob.kjell@ki.se</i><br /><searchLink fieldCode="AR" term="%22Wigerblad%2C+Gustaf%22">Wigerblad, Gustaf</searchLink><relatesTo>2</relatesTo><i> gustaf.wigerblad@ki.se</i><br /><searchLink fieldCode="AR" term="%22Olson%2C+Lars%22">Olson, Lars</searchLink><relatesTo>1</relatesTo><i> lars.olson@ki.se</i><br /><searchLink fieldCode="AR" term="%22Svensson%2C+Camilla+I%2E%22">Svensson, Camilla I.</searchLink><relatesTo>2</relatesTo><i> Camilla.svensson@ki.se</i>
– Name: TitleSource
  Label: Source
  Group: Src
  Data: <searchLink fieldCode="JN" term="%22Journal+of+Neuroscience+Methods%22">Journal of Neuroscience Methods</searchLink>. Apr2011, Vol. 197 Issue 1, p118-127. 10p.
– Name: Subject
  Label: Subjects
  Group: Su
  Data: <searchLink fieldCode="DE" term="%22Neural+development%22">Neural development</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+culture%22">Cell culture</searchLink><br /><searchLink fieldCode="DE" term="%22Astrocytes%22">Astrocytes</searchLink><br /><searchLink fieldCode="DE" term="%22Cellular+signal+transduction%22">Cellular signal transduction</searchLink><br /><searchLink fieldCode="DE" term="%22Laboratory+rats%22">Laboratory rats</searchLink><br /><searchLink fieldCode="DE" term="%22Gene+expression%22">Gene expression</searchLink><br /><searchLink fieldCode="DE" term="%22Spinal+cord+injuries%22">Spinal cord injuries</searchLink><br /><searchLink fieldCode="DE" term="%22Glutamine+synthetase%22">Glutamine synthetase</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Abstract: Primary astrocyte cell cultures have become a valuable tool for studies of signaling pathways that regulate astrocyte physiology, reactivity, and function; however, differences in culture preparation affect data reproducibility. The aim of this work was to define optimal conditions for obtaining primary astrocytes from adult rat spinal cord with an expression profile most similar to adult human spinal cord astrocytes. Hence, we examined whether different Sprague–Dawley substrains and culture conditions affect astrocyte culture quality. Medium supplemented with fetal bovine serum from three sources (Sigma, Gibco, Hyclone) or a medium with defined composition (AM medium) was used to culture astrocytes isolated from spinal cords of adult Harlan and Charles River Spraque–Dawley rats. Purity was significantly different between cultures established in media with different sera. No microglia were detected in AM or Hyclone cultures. Gene expression was also affected, with AM cultures expressing the highest level of glutamine synthetase, connexin-43, and glutamate transporter-1. Interestingly, cell response to starvation was substrain dependent. Charles River-derived cultures responded the least, while astrocytes derived from Harlan rats showed a greater decrease in Gfap and glutamine synthetase, suggesting a more quiescent phenotype. Human and Harlan astrocytes cultured in AM media responded similarly to starvation. Taken together, this study shows that rat substrain and growth medium composition affect purity, expression profile and response to starvation of primary astrocytes suggesting that cultures of Harlan rats in AM media have optimal astrocyte characteristics, purity, and similarity to human astrocytes. [Copyright &y& Elsevier]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Journal of Neuroscience Methods is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=egs&AN=59772851
RecordInfo BibRecord:
  BibEntity:
    Identifiers:
      – Type: doi
        Value: 10.1016/j.jneumeth.2011.02.011
    Languages:
      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        PageCount: 10
        StartPage: 118
    Subjects:
      – SubjectFull: Neural development
        Type: general
      – SubjectFull: Cell culture
        Type: general
      – SubjectFull: Astrocytes
        Type: general
      – SubjectFull: Cellular signal transduction
        Type: general
      – SubjectFull: Laboratory rats
        Type: general
      – SubjectFull: Gene expression
        Type: general
      – SubjectFull: Spinal cord injuries
        Type: general
      – SubjectFull: Glutamine synthetase
        Type: general
    Titles:
      – TitleFull: Influence of rat substrain and growth conditions on the characteristics of primary cultures of adult rat spinal cord astrocytes
        Type: main
  BibRelationships:
    HasContributorRelationships:
      – PersonEntity:
          Name:
            NameFull: Codeluppi, Simone
      – PersonEntity:
          Name:
            NameFull: Gregory, Ebba Norsted
      – PersonEntity:
          Name:
            NameFull: Kjell, Jacob
      – PersonEntity:
          Name:
            NameFull: Wigerblad, Gustaf
      – PersonEntity:
          Name:
            NameFull: Olson, Lars
      – PersonEntity:
          Name:
            NameFull: Svensson, Camilla I.
    IsPartOfRelationships:
      – BibEntity:
          Dates:
            – D: 15
              M: 04
              Text: Apr2011
              Type: published
              Y: 2011
          Identifiers:
            – Type: issn-print
              Value: 01650270
          Numbering:
            – Type: volume
              Value: 197
            – Type: issue
              Value: 1
          Titles:
            – TitleFull: Journal of Neuroscience Methods
              Type: main
ResultId 1