Cellular biology
Our expertise in the development of 2D and 3D human primary cell-based assays has provided data supporting a range of different therapeutic indications from diabetes to fibrosis and oncology, with a significant focus on anti-inflammatory and autoimmune research.
With 30 years’ experience working with a broad range of human primary cell types, our customised approach develops fit-for-purpose assays designed to meet the specific needs of your program. In addition to assay development, our breadth of analytical bioassay platforms enables us to deliver a comprehensive range of assay endpoints, including mediator/cytokine release, changes in gene expression, immunocytochemistry (ICC) , and high content cellular imaging.
Example cell types used for assay development
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Blood cells: PBMCs, monocytes, lymphocytes, neutrophils, monocyte-derived macrophages
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Skin cells: Keratinocytes, fibroblasts
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Lung cells: Bronchial epithelial cells, alveolar macrophages, alveolar type II cells, smooth muscle cells, tumor epithelial cells, cancer-associated fibroblasts
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Liver cells: Hepatocytes, Kupffer cells, stellate cells
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Other cell types: Pancreatic islets, synovial fibroblasts
Culture Formats
2D
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Multiwell plates
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Transwells
3D
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Air-liquid interface
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Matrix-based and matrix free
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Simple or co-culture
Cell-based bioassay endpoints
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Cytokine-mediator release (ELISA and Jess)
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Cell viability and proliferation
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Apoptosis
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Changes in gene expression (qRT-PCR)
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Enzyme activity/release
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Barrier permeability
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Cell migration
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ICC / mIF / Proximity ligation assay
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High content cellular imaging
Cell type(s) | Assay examples |
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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 and 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) |
Lung epithelial organotypic culture model
Organotypic culture of primary human bronchial epithelial cells (hBEC) at air-liquid interface (ALI) culture provides a functional model of the human airway epithelium to enable the study of chronic respiratory disease and delivery of inhaled therapies. Pharmagene® has many years of experience in developing and working with these models, derived from both non-diseased and diseased hBECs, for a variety of applications. In particular ALI cultures have proven an invaluable tool for us to study.
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Mucociliary differentiation
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Mechanisms of mucus secretion, enhanced by stimulated goblet cell hyperplasia
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Epithelial ion transport
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Development of squamous cell metaplasia
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Mucociliary transport
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Drug delivery to the lung
Liver fibrosis organotypic culture model
Using our proprietary culture platform, OrganDOT, we have developed a human liver model for the study of liver fibrosis and screening novel anti-fibrotic therapeutics.
Key features of our Liver Fibrosis OrganDOT model:
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Human primary multicellular, organotypic model (hepatocytes, Kupffers and stellates)
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Viable for up to 4 weeks in culture
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Established stimulus / insult that is common to fibrogenesis
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Characterised changes in fibrotic phenotype at the gene, protein and functional level
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Clinically relevant markers / endpoints