By Evercyte GmbH
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Evercyte human cells as in vitro test systems
Evercyte’s highly differentiated human cells are becoming ever more important as in vitro test systems in various fields of biomedicine, toxicology and biotechnology.
Evercyte possesses an extensive and constantly growing panel of cell types derived from all main relevant human tissues and bodily fluids that can be applied to highly targeted and precise assays in applications that include cytotoxicity testing, cytokine targeting, senolytic and senomorphic drug testing, checking for anti-fibrotic activity or neo-angiogenic potential, and testing wound-healing activity.
In vitro advantages
Although normal somatic cells maintain a phenotype similar to the corresponding cells in vivo, the finite life spans of normal cells limits their application to testing. And while tumor cells can be grown indefinitely, they lose essential characteristics. These disadvantages of normal and tumor cell lines mean most in vitro bioassays cannot be reproduced and standardized.
In order to circumvent these limitations, Evercyte has focused on the establishment of cell lines that don’t suffer from growth arrest while maintaining a phenotype that closely matches the corresponding primary or ‘normal’ cells. It achieves this by ectopic expression of the catalytic subunit of human telomerase in primary cells followed by a detailed characterization of the cellular phenotype.
Cell testing applications
Immortalized human cell types can be applied across a wide range of specific applications that include:
- Cytotoxicity testing: The use of standardized cell types in testing for drug-induced toxicity as well as the potential of drugs to boost cellular proliferation. Using relevant and standardizable human telomerized cell lines, Evercyte offers services for testing drugs for a potential cytotoxic activity, whereby several methods are applied to determine the cell death mechanism, ranking and screening compounds for further testing.
- Cytokine targeting: The search for compounds that target pro-inflammatory mediators is of ever-increasing importance to treat various diseases such osteoarthritis or psoriasis. Using relevant and standardizable human telomerized cell lines, Evercyte offers services for testing compounds for a potency to reduce the release of certain pro-inflammatory cytokines. These assays use telomerized cells treated with compounds followed by measurement of cytokine release using different methods such as western blotting, ELISA or Luminex.
- Senolytic/senomorphic drug testing: Evercyte´s senolytic efficacy and toxicity assays are based on human cells from different tissues and organs (e.g. skin fibroblasts, lung fibroblasts, umbilical vein endothelial cells, microvascular endothelial cells, renal proximal tubular epithelial cells, and mesenchymal stromal cells). As well as growing cells until they enter replicative senescence, Evercyte has established a series of tests for monitoring the cellular phenotype such as monitoring the cell morphology, testing for SA-ß-galactosidase activity, analysis of expression of typical senescence markers (e.g. p16, p21), analysis of cell cycle progression by testing BrdU incorporation, analysis of changes in the senescence associated secretory phenotype (SASP) by measuring secretion of e.g. IL-6, IL-8, MCP-1 in cell culture supernatants or miRNA profiling.
- Anti-fibrotic activity: Evercyte has established relevant and standardizable telomerized fibroblast cell lines, including dermal skin fibroblasts fHDF/TERT166 and lung fibroblasts LF/TERT309, that can be differentiated towards myofibroblasts to provide relevant and standardizable in vitro model systems for fibrosis that will drive the development of new therapeutic approaches.
- Neo-angiogenic potential: Evercyte’s telomerized human umbilical vein endothelial cells (HUVEC/TERT2) form relevant and standardizable in vitro model systems that can mimic neo-angiogenesis in vivo to drive the search for novel drugs.
- Wound-healing activity: Evercyte offers services to test cell migration into a physical gap and test wound-healing activities of certain compounds. It has generated telomerized human fibroblast cells (e.g. fHDF/TERT166 from foreskin, HDF/TERT164 from adult skin, LF/TERT309 from lung) that can be grown without limitation while maintaining a primary-like phenotype and can be seeded into chamber slides to monitor the process of cell migration.
All of these testing services deliver results in between two and six weeks, depending on number of samples or compounds to be tested, and number of replicates and concentrations requested.
Evercyte cell catalogs
Evercyte’s cell banks are categorized by the human tissue or body fluid from which they were derived, including adipose tissue, bone marrow, colon, cornea, kidney, lung, mammary gland, muscle, placenta, skin, thymus, umbilical cord, urine, vascular system, with endometrium cell lines in advanced development.
Evercyte also catalogs cells by cellular type, principally connective tissue cells, epithelial cells, endothelial cells, mesenchymal stromal cells, and muscle cells.
Testing senolytic activity of compounds: Human quiescent and senescent (SIPS) skin fibroblasts are treated with compounds and cellular viability is measured after 3 days. This allows Evercyte to identify compounds with a potential senolytic (blue) activity against senescent cells as well as cytotoxic activity (gray) against growth arrested control cells.