title: Isoflurane decreases self-renewal capacity of rat cultured neural stem cells.
abstract: BACKGROUND
  In models, isoflurane produces neural and behavioral deficits in vitro and in vivo. This study tested the hypothesis that neural stem cells are adversely affected by isoflurane such that it inhibits proliferation and kills these cells.
  
  
  METHODS
  Sprague-Dawley rat embryonic neural stem cells were plated onto 96-well plates and treated with isoflurane, 0.7, 1.4, or 2.8%, in 21% oxygen for 6 h and fixed either at the end of treatment or 6 or 24 h later. Control plates received 21% oxygen under identical conditions. Cell proliferation was assessed immunocytochemically using 5-ethynyl-2'-deoxyuridine incorporation and death by propidium iodide staining, lactate dehydrogenase release, and nuclear expression of cleaved caspase 3. Data were analyzed at each concentration using an ANOVA; P < 0.05 was considered significant.
  
  
  RESULTS
  Isoflurane did not kill neural stem cells by any measure at any time. Isoflurane, 1.4 and 2.8%, reduced cell proliferation based upon 5-ethynyl-2'-deoxyuridine incorporation, whereas isoflurane, 0.7%, had no effect. At 24 h after treatment, the net effect was a 20-30% decrease in the number of cells in culture.
  
  
  CONCLUSIONS
  Isoflurane does not kill neural stem cells in vitro. At concentrations at and above the minimum alveolar concentrations required for general anesthesia (1.4 and 2.8%), isoflurane inhibits proliferation of these cells but has no such effect at a subminimum alveolar concentration (0.7%). These data imply that dosages of isoflurane at and above minimum alveolar concentrations may reduce the pool of neural stem cells in vivo but that lower dosages may be devoid of such effects.
authors.affiliation: Department of Anesthesia, Harvard Medical School, Brigham & Women's Hospital, Boston, Massachusetts 02115, USA. dculley@zeus.bwh.harvard.edu
authors.forename: Deborah J
authors.initials: DJ
authors.lastname: Culley
authors.forename: Justin D
authors.initials: JD
authors.lastname: Boyd
authors.forename: Arvind
authors.initials: A
authors.lastname: Palanisamy
authors.forename: Zhongcong
authors.initials: Z
authors.lastname: Xie
authors.forename: Koji
authors.initials: K
authors.lastname: Kojima
authors.forename: Charles A
authors.initials: CA
authors.lastname: Vacanti
authors.forename: Rudolph E
authors.initials: RE
authors.lastname: Tanzi
authors.forename: Gregory
authors.initials: G
authors.lastname: Crosby
mesh_terms: D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D002273:Carcinogens / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D003470:Culture Media; D003857:Deoxyuridine / Q000031:analogs & derivatives; D005260:Female; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007530:Isoflurane / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D058953:Neural Stem Cells / Q000187:drug effects*; D011247:Pregnancy; D011419:Propidium; D051381:Rats; D017207:Rats, Sprague-Dawley; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology
chemical_list: D018685:Anesthetics, Inhalation; D002273:Carcinogens; D004396:Coloring Agents; D003470:Culture Media; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D055748:SOXB1 Transcription Factors; C527784:Sox2 protein, rat; D011419:Propidium; D007530:Isoflurane; D007770:L-Lactate Dehydrogenase; C031086:5-ethynyl-2'-deoxyuridine; D019311:Staurosporine; D003857:Deoxyuridine