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Neutral red uptake for starting doses for acute oral toxicity

Topic : Acute toxicity, Basal Cytotoxicity

Test Method Number: TM2003-01 (US)
Short Name of TM: NRU cytotoxicity
Year received: 2003
Responsible Organisation: ICCVAM/NICEATM - United States '
Protocol(s)/SOP(s): 139
General Comments:

The NICEATM/ECVAM Validation finalised/OECD Guidance Document

Succeeding TM: TM2007-03 - 3T3 Neutral Red Uptake cytotoxicity assay

Method Description

The in vitro Neutral Red Uptake (NRU) test method is a cell-based approach that assesses the cellular uptake of a dye in the presence of a particular substance. The protocols use normal human epidermal keratinocytes (NHK) and BALB/c 3T3 mouse fibroblasts (3T3). NHK cells are primary cells while 3T3 cells are an inmortal line.

The method is used to assess basal cytotoxicity quantified through IC50 values (the concentration of test substance needed to decrease cell viability by 50% in the cell culture) by spectrophotometric measurments of the cellular uptake of NR dye.

Assays that assess this type of toxicity look at how chemicals exert effects by disrupting structures and functions that are universal to all cells (e.g. cell membrane integrity, energy production and metabolism, transportation of molecules, secretion of molecules).This is opposed to assays that assess the effects of chemicals on specific molecular targets (such as...

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Track Approval Status

  • Submission
  • Validation
  • Peer-review
  • Recommendation
  • Regulatory acceptance/Standards
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