Continuous growth of telomerase-immortalised fibroblasts: How long do cells remain normal?
Open accessAbstract
Previously, we reported successful immortalisation following hTERT introduction in primary human fibroblasts, strain VH25. Since one subclone in that study developed some abnormalities, we decided to study eight additional independent immortalised clones to get an indication of the frequency and type of abnormalities that develop after hTERT-mediated immortalisation. We show that although some cell lines can maintain a normal phenotype for 500 population doublings (PDs), in four clones after 150-300PDs changes developed in basal and radiation-induced p53 and p21(WAF-1,CIP-1) levels. Our experiments demonstrate that, after prolonged culture, cells with abnormalities in cell cycle control parameters can take over the population. This calls for caution when working with hTERT-immortalised cells in vitro as well as in vivo.
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Continued in vitro proliferation under ambient oxygen can itself be mutagenic.
“Although some reports claim that hTERT-immortalization is nonmutagenic and maintains diploidy during extended culture (32, 33), our observations, and those of others (34 -36), suggest instead that continued in vitro proliferation under ambient oxygen can itself be mutagenic.”
From Why Cockayne syndrome patients do not get cancer despite their DNA repair deficiency (2016) · cites this paper as [35]
The paper reports that four of eight hTERT-immortalised fibroblast clones developed abnormal cell-cycle responses or ploidy after prolonged culture. It also suggests that DNA damage during culture may lead to genomic instability. The passages do not specify ambient oxygen as the condition responsible or directly demonstrate that continued proliferation itself caused mutations. (LLM, conf=0.84)
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Cites
Extension of Life-Span by Introduction of Telomerase into Normal Human Cells
Bodnar, A. G., Ouellette, M., Frolkis, M., Holt, S. E., Chiu, C.-P., Morin, G. B., Harley, C. B., Shay, J. W., Lichtsteiner, S., & Wright, W. E. (1998) · Science · doi:10.1126/science.279.5349.349 · Cited by 4,975 · see citing works
Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors
Campisi, J. (2005) · Cell · doi:10.1016/j.cell.2005.02.003 · Cited by 2,359 · see citing works
The signals and pathways activating cellular senescence
Ben-Porath, I., & Weinberg, R. A. (2005) · The International Journal of Biochemistry & Cell Biology · doi:10.1016/j.biocel.2004.10.013 · Cited by 1,046 · see citing works
Absence of cancer–associated changes in human fibroblasts immortalized with telomerase
Morales, C. P., Holt, S. E., Ouellette, M. M., Kaur, K. J., Yan, Y., Wilson, K. S., White, M. A., Wright, W. E., & Shay, J. W. (1999) · Nature Genetics · doi:10.1038/5063 · Cited by 805 · see citing works
Jiang, X., Jimenez, G. S., Chang, E., Frolkis, M., Kusler, B., Sage, M., Beeche, M., Bodnar, A. G., Wahl, G. M., Tlsty, T. D., et al. (1999) · Nature Genetics · doi:10.1038/5056 · Cited by 668 · see citing works
p53 mutation heterogeneity in cancer
Soussi, T., & Lozano, G. (2005) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2005.03.190 · Cited by 275 · see citing works
(2003) · PubMed · Cited by 146 · see citing works
Alterations in the p16INK4a and p53 tumor suppressor genes of hTERT-immortalized human fibroblasts
Noble, J. R., Zhong, Z.-H., Neumann, A. A., Melki, J. R., Clark, S. J., & Reddel, R. R. (2004) · Oncogene · doi:10.1038/sj.onc.1207440 · Cited by 53 · see citing works
Nakamura, H., Fukami, H., Hayashi, Y., Kiyono, T., Nakatsugawa, S., Hamaguchi, M., & Ishizaki, K. (2002) · Journal of Radiation Research · doi:10.1269/jrr.43.167 · Cited by 35 · see citing works
Kampinga, H. H., van Waarde-Verhagen, M. A. W. H., Assen‐Bolt, A. J. V., Nieuwenhuis, B., Rodemann, H. P., Prowse, K. R., & Linskens, M. H. K. (2004) · International Journal of Radiation Biology · doi:10.1080/09553000410001692735 · Cited by 21 · see citing works
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Flagged first, then by citations
Cited by
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Telomerase, senescence and ageing
Shawi, M., & Autexier, C. (2008) · Mechanisms of Ageing and Development · doi:10.1016/j.mad.2007.11.007 · Cited by 156 · see citing works
“…phenotype, although there are two reports describing cell-cycle abnormalities in fibroblasts and neoplastic transformation of mesenchymal stem cells (Jiang et al., 1999; Klinger et al., 2006; Moon and Jarstfer, 2007; Morales et al., 1999; Shay and Wright, 2005; van Waarde-Verhagen et al., 2006).”
Wright, L. S., Prowse, K. R., Wallace, K., Linskens, M. H. K., & Svendsen, C. N. (2006) · Experimental Cell Research · doi:10.1016/j.yexcr.2006.03.012 · Cited by 134 · see citing works
“Furthermore, recent data indicate that other hTERT immortalized somatic cell lines show changes in cell cycle regulatory genes and karyotypes after prolonged culturing [47 – 50].”
BackgroundSelf-citationWhy Cockayne syndrome patients do not get cancer despite their DNA repair deficiency
Reid-Bayliss, K. S., Arron, S. T., Loeb, L. A., Bezrookove, V., & Cleaver, J. E. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1610020113 · Cited by 55 · see citing works
Hung, C. L.-K., Maiuri, T., Bowie, L. E., Gotesman, R. D., Son, S., Falcone, M., Giordano, J. V., Gillis, T., Mattis, V. B., Lau, T. C., et al. (2018) · Molecular Biology of the Cell · doi:10.1091/mbc.e18-09-0590 · Cited by 40 · see citing works
Impact on oxidative phosphorylation of immortalization with the telomerase gene
Auré, K., Mamchaoui, K., Frachon, P., Butler-Browne, G. S., Lombès, A., & Mouly, V. (2007) · Neuromuscular Disorders · doi:10.1016/j.nmd.2007.01.019 · Cited by 31 · see citing works
“However the number of divisions gained should be high, above 150–300 divisions [26].”
BackgroundSynergistic induction of profibrotic PAI-1 by TGF-β and radiation depends on p53
Niemantsverdriet, M., de Jong, E., Langendijk, J. A., Kampinga, H. H., & Coppes, R. P. (2010) · Radiotherapy and Oncology · doi:10.1016/j.radonc.2010.04.002 · Cited by 16 · see citing works
Self-citationGrzes, M., Jaiswar, A., Grochowski, M., Wojtyś, W., Kaźmierczak, W., Olesiński, T., Lenarcik, M., Nowak-Niezgoda, M., Kołos, M., Canarutto, G., et al. (2024) · Cell Death and Disease · doi:10.1038/s41419-024-06965-3 · Cited by 15 · see citing works
“Human h-TERT (telomerase reverse transcriptase) immortalized K15 and K21 fi broblasts were a kind gift from Prof. Harm Kampinga (University of Groningen, Netherlands) [20].”
Biela, A. P., Coste, F., Culard, F., Guérin, M., Goffinont, S., Gasteiger, K. L., Cieśla, J., Winczura, A., Kazimierczuk, Z., Gasparutto, D., et al. (2014) · Nucleic Acids Research · doi:10.1093/nar/gku613 · Cited by 14 · see citing works
LAMP2A-mediated autophagy involved in Huntington’s disease progression
Choi, S. H., & Cho, K. (2020) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2020.11.042 · Cited by 12 · see citing works
Cytogenetic features of human trophoblast cell lines SWAN-71 and 3A-subE.
Reiter, J., Drendel, H. M., Chakraborty, S., Schellinger, M., Lee, M., & Mor, G. (2017) · Placenta · doi:10.1016/j.placenta.2017.02.010 · Cited by 9 · see citing works
Immediate and gradual gene expression changes in telomerase over-expressing fibroblasts
Daniels, D. J., Clothier, C., Swan, D., & Saretzki, G. C. (2010) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2010.07.018 · Cited by 7 · see citing works
Ohshima, S., & Seyama, A. (2016) · Genes Chromosomes and Cancer · doi:10.1002/gcc.22354 · Cited by 4 · see citing works
“…results in transformed phenotypes such as loss of contact inhibition (Milyavsky et al., 2003), karyotypic changes (Mondello et al., 2003), anchorage-independent growth (Zongaro et al., 2005), and growth acceleration in prolonged culture (Milyavsky et al., 2003; van Waarde-Verhagen et al., 2006).”
BackgroundGrzes, M., Jaiswar, A., Grochowski, M., Wojtyś, W., Kaźmierczak, W., Olesiński, T., Lenarcik, M., Nowak-Niezgoda, M., Kołos, M., Canarutto, G., et al. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.12.20.572548 · Cited by 1 · see citing works
“Human h-TERT immortalized K15 fibroblast were a kind gift from Prof. Harm Kampinga (Univ. of Groningen) 17 .”
MethodGrzes, M., Jaiswar, A., Kaźmierczak, W., Olesiński, T., Nowak-Niezgoda, M., & Walerych, D. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.09.04.749381
- Authors
- van Waarde-Verhagen, M. A. W. H., Kampinga, H. H., & Linskens, M. H. K.
- Affiliations
- University Medical Center Groningen (NL)
- University of Groningen (NL)
- Corresponding author
- Linskens, M.H.K.
- ORCID
- Kampinga, H.H.
- Venue
- Mechanisms of Ageing and Development
- Publisher
- Elsevier BV
- Published
- 2006-01
- Volume
- 127
- Issue
- 1
- Pages
- 85-87
- ISSN
- 0047-6374 (ISSN-L)
- OpenAlex
- W2068286052
- Cited by
- 14
- References
- 10
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 1 · 2020: 1 · 2018: 1 · 2017: 1 · 2016: 2 · 2014: 1
- FWCI
- 0.60
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Mechanisms of Ageing and Development (publishedVersion)
- PubMed (publishedVersion)
- University of Groningen research database (University of Groningen / Centre for Information Technology) (submittedVersion, other-oa)
- Funders
- Ministry of Earth Sciences
- Topics
- Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Keywords
- telomerase, hTERT, human fibroblasts, cell immortalization, p53, p21, cell cycle regulation, population doubling
- MeSH terms
- Cell Cycle, Cells, Cultured, Fibroblasts, Humans, Tumor Suppressor Protein p53, Cellular Senescence, Telomerase, Cyclin-Dependent Kinase Inhibitor p16, Cell Proliferation, Cyclin-Dependent Kinase Inhibitor p21
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