Isoflurane Decreases Self-renewal Capacity of Rat Cultured Neural Stem Cells

• Published in: Anesthesiology (Philadelphia) 2011-10, Vol.115 (4), p.754-763
• Main Authors: CULLEY, Deborah J, BOYD, Justin D, PALANISAMY, Arvind, ZHONGCONG XIE, KOJIMA, Koji, VACANTI, Charles A, TANZI, Rudolph E, CROSBY, Gregory
• Format: Article
• Language: English
• Subjects: Anesthesia; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Anesthetics, Inhalation - pharmacology; Animals; Biological and medical sciences; Carcinogens - pharmacology; Cell Death - drug effects; Cell Proliferation - drug effects; Cells, Cultured; Coloring Agents; Culture Media; Deoxyuridine - analogs & derivatives; Female; Image Processing, Computer-Assisted; Immunohistochemistry; Intermediate Filament Proteins - metabolism; Isoflurane - pharmacology; L-Lactate Dehydrogenase - metabolism; Medical sciences; Nerve Tissue Proteins - metabolism; Nestin; Neural Stem Cells - drug effects; Pregnancy; Propidium; Rats; Rats, Sprague-Dawley; SOXB1 Transcription Factors - metabolism; Staurosporine - pharmacology
• ISSN: 0003-3022; 1528-1175
• DOI: 10.1097/aln.0b013e318223b78b

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. 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. 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. 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.