Services

cell line characterization

Inherent variation of increasingly complex in vitro test systems as well as little guidance for routine cell culture reduce the predictive value of cell cultures. Therefore, setting common standards is of ever-increasing importance in view to the high need for comparability and reproducibility of research and development in life sciences. In this regard the characterization of the cellular phenotype for presence of marker proteins or functional properties is of utmost importance. Additionally, cells need to be tested for presence of contaminations and authentication.

General information

Testing functional properties: Evercyte offers services for characterization of the cellular phenotype and functional properties; services offered strongly depend on the cells of interest (e.g. staining for presence of marker proteins, testing of differentiation properties, response to hormones or cytokines, cell migration, tumorigenicity tests, NGS)  

Testing for contaminations: services offered include e.g. sterility testing (testing for presence of bacteria incl. mycoplasma and fungi), testing for presence of human pathogenic viruses (e.g. HIV, HepA, HepB, HepC, Parvo B19), testing for cross-contaminations / authentication using STR profiling
Testing in vitro growth properties:
Evercyte offers services for testing long term growth properties in vitro  (growth of cells up to at least 50 population doublings, establishment of growth curves) and testing different cell culture media compositions (proliferation and differentiation media)

Example – characterization of cellular functions

neo-angiogenic properties of endothelial cells  

Human umbilical vein endothelial cells HUVEC/TERT2 are characterized for neo-angiogenic properties, which are key characteristics of endothelial cells.

When seeded onto matrigel matrix HUVEC/TERT2 cells start formation of tubule-like structures (left). Additionally, HUVEC/TERT2 cells can be grown as 3D spheroids embedded in a collagen matrix. Treatment of these spheroids with vascular endothelial growth factor (VEGF) leads to formation of sprouts (right). These data demonstrate that telomerized endothelial cells, albeit growing without limitations, show essential differentiated properties and neo-angiogenic potential.

Example – characterization of cellular functions

induction of myofibroblast differentiation

Treatment of skin fibroblasts fHDF/TERT166 cells with transforming growth factor ß (TGFß) induces expression of alpha smooth muscle actin (alpha SMA) as demonstrated by immunofluorescent staining (green: alpha SMA, blue: cell nuclei stained with DAPI). These data show the cells ability to differentiate towards myofibroblasts, which is a key event in physiological and pathological tissue repair.

Example - characterization of cellular phenotype

testing the tumorigenic potential of cells using soft agar assay

Cells are seeded at different densities into soft agar and cellular proliferation and colony formation is measured using a fluorescent, nucleic acid binding dye. Tumorigenic human embryonic kidney cells HEK293 are used as positive control for anchorage independent growth. Whereas, HEK293 cells form colonies in soft agar (left picture), telomerized mesenchymal stem cells (MSC/TERT) and the corresponding normal cells (MSC) do not proliferate when grown in soft agar and do not aquire a tumorigenic phenotype.

Leaflets - telomerized cells

Adipose-derived mesenchymal stem cells ASC/TERT1
Lung epithelial cells HBEC3-KT
Colon epithelial cells HCEC-1CT
Corneal epithelial cells hTCEpi
Skin keratinocytes NHEK/SVTERT3-5
Podocytes PODO/TERT256
Renal proximal tubular epithelial cells RPTEC/TERT1
Myoblast cells LHCN-M2
Wharton´s Jelly derived mesenchymal stem cells WJ-MSC/TERT273
Umbilical vein endothelial cells HUVEC/TERT2

 

Selected publications


Zhou T, Benda C, Duzinger S, Huang Y, Li X, Li Y, Guo X, Cao G, Chen S, Hao L, Chan YC, Ng KM, Ho JC, Wieser M, Wu J, Redl H, Tse HF, Grillari J, Grillari-Voglauer R, Pei D, Esteban MA (2011) Generation of induced pluripotent stem cells from urine. J Am Soc Nephrol. 2011 Jul;22(7):1221-8. https://pubmed.ncbi.nlm.nih.gov/33717998/
Benda C, Zhou T, Wang X, Tian W, Grillari J, Tse HF, Grillari-Voglauer R, Pei D, Esteban MA. Urine as a source of stem cells. Adv Biochem Eng Biotechnol. 2013;129:19-32. https://pubmed.ncbi.nlm.nih.gov/23038280/
Zhou T, Benda C, Dunzinger S, Huang Y, Ho JC, Yang J, Wang Y, Zhang Y, Zhuang Q, Li Y, Bao X, Tse HF, Grillari J, Grillari-Voglauer R, Pei D, Esteban MA. Generation of human induced pluripotent stem cells from urine samples. Nat Protoc. 2012 Dec;/(12):2080-9. https://pubmed.ncbi.nlm.nih.gov/23138349/
Chang MW, Grillari J, Mayrhofer C, Fortschegger K, Allmaier G, Marzban G, Katinger H, Voglauer R. Comarison of early passage, senescent and hTERT immortalized endothelial cells. Exp Cell Res. 2005 Sept;309(1):121-36. https://pubmed.ncbi.nlm.nih.gov/15964568/
Wolbank S, Stadler G, Peterbauer A, Gillich A, Karbiener M, Streubel B, Wieser M, Katinger H, van Griensven M, Redl H, Gabriel C, Grillari J, Grillari-Voglauer R. Telomerase immortalized human amnion-derived mesenchymal stem cells: maintenance of differentiation and immunomodulatory characteristics. Tissue Eng Part A. 2009 Jul;15(7):1843-54. https://pubmed.ncbi.nlm.nih.gov/19125642/
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“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

"Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed."

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

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Customer Reviews

“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

"Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed."

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

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