In Vitro Models

LRRI offers a number of in vitro assays for determining the pharmacology and toxicology of pharmaceuticals, toxicants, and mixtures. We place particular emphasis on assays that can be used to assess the impact on the respiratory tract. Cell cultures use both primary and transformed cell lines originating from a number of primary cell types. Specialized capabilities include air-liquid interface systems, isolated perfused lung, and mucociliary functional assays. When appropriate, experiments can be conducted using Good Laboratory Practices.

Key Capabilities:
  • Cell culture: primary cells, transformed cells
  • Isolation of primary cells
  • Cytotoxicity assays: lactate dehydrogenase, trypan blue, tetrazolium conversion assays
  • Air-liquid interface exposures of cells to aerosols (diesel exhaust, cigarette smoke, toxins, drugs)
  • Isolated perfused lung (metabolism, aerosol exposure, PK, etc.)
  • Cytokine production and secretion
  • Western blots and ELISA for protein, various targets
  • Macrophage phagocytosis assays
  • Peripheral blood mononuclear cell stimulation
  • Mast cell degranulation
  • Cytochrome P-450 activity assays
  • In vitro microsomal metabolism, metabolite identification
  • Glutathione (oxidized, reduced) measurements
  • Lipid peroxidation measurements
  • Protein carbonyl measurements
  • Gene expression by RT-PCR or qRT-PCR
  • Zymography for metalloproteinase activity
  • Infection of primary cells with adenoviral expression vectors
  • Luciferase promoter response assays
  • Mucociliary functional assays on differentiated primary tracheal epithelial cells

Virology: Multiple clinical and Select Agent viral strains. Agents include: Respiratory syncytial virus, influenza (H3 and H1 to include newest H1N1 “swine flu”), highly pathogenic avian, adenovirus, metapneumovirus, hantavirus, SARS coronavirus, monkeypox virus.

Bacteriology: Multiple clinical and Select Agent bacterial strains. Bacterial and mammalian co-cultures to test infectivity and cytotoxicity. Agents include: Pseudomonas aeruginosa, Group B Streptococcus, Staphylococcus aureus, Klebsiella pneumoniae, Bacillus sp., F. tularenis (LVS strain), F. tularensis (Shu4 strain), B. anthracis (Ames strain), Y. pestis (Colorado 92 strain).

AAALAC, FDA, USDA, state, and other regulatory licensure in hand