Variation in Pentose Phosphate Pathway-Associated Metabolism Dictates Cytotoxicity Outcomes Determined by Tetrazolium Reduction Assays

Details

• Personal Author:
  Coyle JP ; Derk R ; Farcas M ; Jensen J ; Johnson C ; Kornberg TG ; Rojanasakul LW ; Rojanasakul Y ; Stueckle, Todd A.
• Description:
  Tetrazolium reduction and resazurin assays are the mainstay of routine in vitro toxicity batteries. However, potentially erroneous characterization of cytotoxicity and cell proliferation can arise if verification of baseline interaction of test article with method employed is neglected. The current investigation aimed to demonstrate how interpretation of results from several standard cytotoxicity and proliferation assays vary in dependence on contributions from the pentose phosphate pathway (PPP). Non-tumorigenic Beas-2B cells were treated with graded concentrations of benzo[a]pyrene (B[a]P) for 24 and 48 h prior to cytotoxicity and proliferation assessment with commonly used MTT, MTS, WST1, and Alamar Blue assays. B[a]P caused enhanced metabolism of each dye assessed despite reductions in mitochondrial membrane potential and was reversed by 6-aminonicotinamide (6AN)-a glucose-6-phosphate dehydrogenase inhibitor. These results demonstrate differential sensitivity of standard cytotoxicity assessments on the PPP, thus (1) decoupling "mitochondrial activity" as an interpretation of cellular formazan and Alamar Blue metabolism, and (2) demonstrating the implicit requirement for investigators to sufficiently verify interaction of these methods in routine cytotoxicity and proliferation characterization. The nuances of method-specific extramitochondrial metabolism must be scrutinized to properly qualify specific endpoints employed, particularly under the circumstances of metabolic reprogramming. [Description provided by NIOSH]
• Subjects:
  Cytotoxins Laboratories Occupational Health Safety Toxicology
• Keywords:
  Cell Culture Techniques Cell Proliferation Cytotoxic Effects Cytotoxicity In Vitro Studies Laboratory Techniques
• ISSN:
  2045-2322
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  Massachusetts ; OSHA Region 1 ; OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  13
• NIOSHTIC Number:
  nn:20067676
• Citation:
  Sci Rep 2023 May; 13:8220
• Contact Point Address:
  Jayme P. Coyle, HELD/ACIB, National Institute for Occupational Safety and Health, Morgantown, WV, USA
• Email:
  jaymec2010@gmail.com
• Federal Fiscal Year:
  2023
• NORA Priority Area:
  Manufacturing
• Peer Reviewed:
  True
• Source Full Name:
  Scientific Reports
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:1e7cf1aaf88f69a6adc2161fa9f3a56589e53fa0c1f0b7f91556be04eaf444a0e26888d5514853335d741f406c758ef2841eac3f002f80fe72bb87b6ad598fba
• Download URL:
  https://stacks.cdc.gov/view/cdc/229944/cdc_229944_DS1.pdf
• File Type:
  [PDF - 4.69 MB ]