Service life span extension2022-02-17T12:45:03+00:00

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life span extension

With in-depth knowledge of cellular senescence and mechanisms to overcome this irreversible growth arrest, Evercyte offers the development of human cell lines that can be grown without limitations while maintaining a phenotype similar to the corresponding normal cells. Besides ectopic expression of the catalytic subunit of human telomerase (hTERT) or regulators of cell cycle control pathways, Evercyte has also established protocols to reprogram cells towards induced pluripotent stem cells (iPSCs) that can give rise to theoretically any cell type of the human organism.

General information

Tissue, cell type: Evercyte offers the immortalization / life span extension of different human cell types (e.g. fibroblasts from skin, lung, kidney tissue, keratinocytes, endothelial cells from skin, umbilical vein or mesenchymal stem cells from adipose tissue, placental tissue or bone marrow)

Ethical issues: cells used for life span extension are isolated with ethical approval from the responsible IRB and prior given written informed consent by the donor
Life span extension method:
depending on the cell type, cells are immortalized by ectopic expression of the catalytic subunit of human telomerase (hTERT) and/or cell cycle regulators such as cyclin-dependent-kinase 4 (cdk-4) or viral oncogenes (e.g. SV40 early region, SV40 largeT/smallt antigens)
Selection of transfectants:
positively transfected cells are selected using antibiotic resistance genes such as neomycin-phosphotransferase (neo)
Characterization:
selected cell lines are characterized depending on the customers needs; Evercyte offers services to test the presence of cell type specific markers and functions, authentication, freedom from contaminations, tumorigenic potential or RNAseq

Recent example – immortalization of mesenchymal stem cells 

Wharton´s Jelly-derived mesenchymal stem cells were isolated from umbilical cord tissue using enzymatic and mechanical disaggregation. After enrichment, the normal cells were transfected with a plasmid carrying the catalytic subunit of human telomerase (hTERT) and transfectants were selected by growing cells in the presence of Geneticin sulfate (G418). The resulting cell line can be grown for a minimum of 50 population doublings with a stable growth rate without showing signs of growth retardation. The cells are characterized by the typical spindle-shaped mesenchymal morphology and express typical markers and functions of mesenchymal stem cells (not shown). 

Recent example – reprogramming of urine cells into iPSCs

Evercyte has developed protocols for isolation of different cells (e.g. renal proximal tubular epithelial cells, podocytes, urine-stem cells, mesenchymal stem cells) from human urine sediments. These cells are reprogrammed into induced pluripotent stem cells (iPSCs) that can be differentiated towards cells from the 3 germ layers such as muscle cells, neuronal cells or hepatocyte-like cells.

FAQs

Are telomerized cells tumorigenic?2021-12-15T09:49:29+00:00

Ectopic expression of the catalytic subunit of human telomerase hTERT does not induce a tumorigenic conversion of the cells. The cells do not form colonies when seeded into soft agar to test anchorage independent growth as a hallmark of tumor cells (in vitro assay) and do not form tumors in nude mice (in vivo assay).

Does overexpression of hTERT change the cellular phenotype?2021-12-15T09:34:44+00:00

Ectopic expression of the catalytic subunit of human telomerase hTERT leads to an extension of the cellular life span while maintaining cell type specific markers and functions. Therefore, the cells are characterized by a phenotype that is similar to that of the corresponding normal cells.

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
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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