Early Safety Screening Cytotoxicity

Early Safety Screening Cytotoxicity in Human Primary Cells

Regulatory authorities are increasingly requiring that predictive cell-based assays be performed to assess the potential toxic effects of developmental compounds. Ricerca Biosciences has developed a reliable, high throughput model of cytotoxicity testing that provides a quick and cost-effective option suitable for early toxicity liability assessment.

 

Cytotoxicity assay

This assay employs high-resolution cell imaging that allows more sensitive detection of cell loss and cell proliferation inhibition. The assay can be run with a single cell type, the pre-validated primary cell panel or cells chosen through collaboration with our expert scientists who have experience running this assay with hundreds of human cell lines.

 

This cost-effective early-phase assay provides rapid and reliable results while saving clients both time and money.

 

Service features

  • Cytotoxicity Screening with High Content Analysis (HCA), the nuclear fluorescent signal is measured using quantitative image analysis
  • Compound testing in pre-validated human primary cells:
    • Catalog Number 970010: Human Primary Hepatocytes (HPH)
    • Catalog Number 970020: Normal Human Dermal Fibroblasts (NHDF-neo)
    • Catalog Number 970030: Human Renal Proximal Tubular Epithelial Cells (HRPTEpiC)
    • Catalog Number 970040: Human Umbilical Vein Endothelial Cells (HUVEC)
    • Catalog Number 970050: Human Umbilical Mesenchymal Stem Cells (HUMSC)
    • Catalog Number 970060: Human primary cell panel
  • Single-plex cell viability assay - relative cell numbers EC50 and IC50
  • 4 compound minimum
  • Test compound evaluation: 10-pt dose response curve up to 100μM, 3.16-fold serial dilution with each concentration performed in triplicate at 24 hr exposure
  • Compound requirements: 200μL of a 20mM solution or equivalent amount in pre-weighed vial
  • Final DMSO concentration: 0.5%
  • Reference compound included as a control on each plate, vehicle: 0.5% DMSO
  • Aqueous solubility options

Assay design and data output

  • Human primary cells harvested from single donor source and purified by quality controlled vendors
  • Reference compound included on every plate
  • Minimum compound requirements
  • Compound incubation: 24 hours
  • Reliable and sensitive high-content cellular image analysis
    • High throughput (384 well)
    • Generates data at 10 concentrations (in triplicates) for DMSO soluble compounds (Aqueous compound solubility also available)
  • Automated fluorescence microscopy carried out using a GE Healthcare INCell Analyzer 1000
  • Images collected with a 4X objective to capture more cells and provide better statistics
  • Relative cell number measured by nuclear segmentation of the incorporated nuclear dye
  • Cell viability output is referred to as the percentage of live cells quantified by nuclear segmentation
    • Nuclear segmentation data output is transformed to percent of control (POC) after normalizing relative cell count data in treated wells to untreated vehicle wells
  • Assay output: Relative cell number EC50/IC50
    • EC50 is a concentration at the curve inflection point or half the effective response
    • IC50 is a concentration at 50% of maximal possible response
  • Curve-fitting, EC50/IC50 calculations and report generation are performed using a custom data reduction engine MathIQ based software (AIM)
  • 4-week turnaround time