Passage Variation of PC12 Cells Results in Inconsistent Susceptibility to Externally Induced Apoptosis

Nihar Kinarivala, Kaushik Shah, Thomas J. Abbruscato, Paul C. Trippier

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

The PC12 cell line is a widely used in vitro model for screening the neuroprotective activity of small molecule libraries. External insult due to serum deprivation or addition of etoposide induces cell death by apoptosis. While this screening method is commonly used in early stage drug discovery no protocol accounting for cell passage number effect on neuroprotective activity has been disclosed. We herein report that passage variation results in false-positive/false-negative identification of neuroprotective compounds; undifferentiated PC12 cells with high passage number are less sensitive to injury induced by serum-deprivation or etoposide treatment. In contrast, NGF differentiated PC12 cells of later passage number are more sensitive to injury induced by etoposide than lower passage number but only after 72 h. Passage number also affects the adherence phenotype of the PC12 cells, complicating screening assays. We report an optimized protocol for screening the neuroprotective activity of small molecules in PC12 cells, which accounts for passage number variations.

Original languageEnglish (US)
Pages (from-to)82-88
Number of pages7
JournalACS Chemical Neuroscience
Volume8
Issue number1
DOIs
StatePublished - Jan 18 2017

Keywords

  • PC12 cells
  • apoptosis
  • cell viability
  • neuroprotection assay
  • protocol
  • screening

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Cognitive Neuroscience
  • Cell Biology

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