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Cell-Based Assays and Imaging

Cell-based assays in both 2D and 3D formats across a broad range of human primary cell types
Our assays have provided data supporting a range of different therapeutic indications from diabetes to fibrosis and oncology, but our primary focus remains in anti-inflammatory and autoimmune research. Our customized approach develops fit-for-purpose assays to the specific needs of your program. In addition to assay development, our breadth of analytical platforms enables us to deliver a comprehensive range of assay endpoints, including mediator/cytokine release, changes in gene expression, ICC, and high content cellular imaging.

Cell-Based assays customized to your program

Example Cell Types Used for Assay Development


  • Blood cells: PBMCs, monocytes, lymphocytes, neutrophils, monocyte-derived macrophages
  • Skin cells:  keratinocytes, fibroblasts
  • Lung cells: bronchial epithelial cells, alveolar macrophages, alveolar type II cells, smooth muscle cells, tumor epithelial cells, cancer-associated fibroblasts
  • Liver cells: hepatocytes, Kupffer cells, stellate cells
  • Other cell types:  pancreatic islets, synovial fibroblasts

Culture Formats


  • 2D
    • multiwell plates
    • transwells
  • 3D
    • air-liquid interface
    • matrix-based and matrix free
    • simple or co-culture

Cell-Based Assay and Endpoint Types


  • Cytokine-mediator release
  • Cell viability/proliferation
  • Apoptosis
  • Changes in gene expression
  • Enzyme activity/release
  • Barrier permeability
  • Cell migration
  • ICC
  • Multiplex immunofluorescence/high content cellular imaging
  • Automated Western blotting (Jess™)

CELL TYPE(S)ASSAY EXAMPLES

Whole blood

Inhibition of LPS stimulated TNFα and IL-6 release

Stimulation (PHA) and inhibition (CsA) of cell proliferation

PBMCs

Inhibition of OKT3 (CD3) stimulated IL-2 and IFNγ release

Inhibition of CD3:CD28 stimulated TNFα release

Monocytes & Monocyte-derived Macrophages

Inhibition of LPS stimulated TNFα release

Lymphocytes

Inhibition of OKT3 (CD3) stimulated IL-2 release

Neutrophils

Transmigration (+HUVEC)

Superoxide release

HUVECs

Inhibition of thrombin stimulated IL-8 and MCP-1 release

Hepatocytes

Inhibition of TNFα stimulated IL-6 and IL-8 release CYP induction (mRNA)

Stellates

TGF-β1-induced changes in fibrogenic gene expression and collagen deposition

Alveolar macrophages

Inhibition of LPS stimulated TNFα release

Keratinocytes

Stimulation (IL-4) and inhibition (CsA) of cell proliferation

Synovial fibroblasts (RA)

Validated assay for screening novel anti-TNFα therapeutic antibodies (biosimilars)

Case study: bioassay validation supporting EMA marketing authorization application

Details:

  • 2D assay
  • Synovial fibroblasts from rheumatoid arthritis donors

Assay development included:

  • Reference standard inhibitor
  • Titration of stimulus
  • Selection of sentinel marker
  • Formulation effect

Assay validation included:

  • Linearity and range
  • Relative potency
  • Parallelism
  • Specificity
  • Precision—intra-/interassay
  • Biosimilar screen

Precision also performed a biosimilar screen comparing relative potency vs Humira.


Results

1268-IL-6-RA-syn-fibs-2

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