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