The DOI matches its Crossref record; 7 of the 7 references it cites were found in a registry or an index.
Characterization of Growth and Differentiation in a Telomerase-Immortalized Human Corneal Epithelial Cell Line
Abstract
PURPOSE: To develop and characterize a telomerase-immortalized human corneal epithelial cell line (hTCEpi) to serve as an in vitro model for studying the molecular mechanisms involved in regulating human corneal epithelial cell differentiation. METHODS: Primary cultures of human corneal epithelial cells were infected with a retroviral vector encoding human telomerase reverse transcriptase (hTERT). Infected hTCEpi cells were selected, cloned, and characterized to identify telomerase activity, proliferative capacity, karyotype, and differentiative potential in routine culture and under consecutive submerged and air-lifted conditions. Cells were evaluated to measure cell cycle kinetics (anti-Ki-67, anti-p16), stratification (phalloidin and anti-ZO-1), and differentiation (anti-K3, anti-BCL-2 and TUNEL labeling). RESULTS: hTCEpi cells exhibited telomerase activity, a normal karyotype and cell cycle kinetics at greater than 240 population doublings, and loss of p16 after passage 10. Air-lifting produced a well stratified epithelium (five to seven cell layers) with apical ZO-1-stained tight junctions. Submersed culture demonstrated increasing expression of stratification markers (K5/K14) with K3-corneal keratin marker expression in long-term, air-lifted culture. Anti-BCL-2 staining showed both nuclear and cytoplasmic localization with loss of nuclear BCL-2 expression in TUNEL-labeled surface epithelial cells. CONCLUSIONS: hTCEpi cells stratify, differentiate, and desquamate similar to normal human corneal epithelium. Further study of the hTCEpi cell line may be valuable in studying the molecular mechanisms regulating corneal epithelial cell differentiation and desquamation.
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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
Self-citationBCL2 protein is topographically restricted in tissues characterized by apoptotic cell death.
Hockenbery, D. M., Zutter, M., Hickey, W., Nahm, M., & Korsmeyer, S. J. (1991) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.88.16.6961 · Cited by 1,252 · cited 1×
“This is supported by other studies reporting BCL-2 restriction to tissues such as stem cells, which are long-lived and display a high proliferative capacity.(38) Supporting this view, moving from limbus to the central cornea and from the basal to the superficial layer, cytoplasmic BCL-2 expression decreases in a gradi…”
Kiyono, T., Foster, S. A., Koop, J. I., McDougall, J. K., Galloway, D. A., & Klingelhutz, A. J. (1998) · Nature · doi:10.1038/23962 · Cited by 1,206
Vaziri, H., & Benchimol, S. (1998) · Current Biology · doi:10.1016/s0960-9822(98)70109-5 · Cited by 1,003 · cited 1×
“Infection of human cell lines with hTERT leads to the activation of telomerase, preventing telomere erosion and subsequent telomere-dependent senescence.(9) Human cell lines immortalized with hTERT exhibit genetic stability, normal contact inhibition, and maintain the capacity to differentiate.”
Putative telomere-independent mechanisms of replicative aging reflect inadequate growth conditions
Ramirez, R. D., Morales, C. P., Herbert, B.-S., Rohde, J. M., Passons, C., Shay, J. W., & Wright, W. E. (2001) · Genes & Development · doi:10.1101/gad.859201 · Cited by 444 · cited 1×
“Alternatively, it has been reported that p16 induction can be bypassed under appropriate culture conditions.(10,31,32) To examine the role of p16 in hTCEpi immortalization, we assessed p16 expression in hTCEpi cells at early, middle, and late passages.”
Self-citationRheinwald, J. G., Hahn, W. C., Ramsey, M. R., Wu, J. Y., Guo, Z., Tsao, H., De Luca, M., Catricalà, C., & O'Toole, K. M. (2002) · Molecular and Cellular Biology · doi:10.1128/mcb.22.14.5157-5172.2002 · Cited by 338
Genetic and Epigenetic Changes in Human Epithelial Cells Immortalized by Telomerase
Farwell, D. G., Shera, K. A., Koop, J. I., Bonnet, G. A., Matthews, C. P., Reuther, G. W., Coltrera, M. D., McDougall, J. K., & Klingelhutz, A. J. (2000) · The American Journal of Pathology · doi:10.1016/s0002-9440(10)65025-0 · Cited by 196
Cited by
Review of hTERT-Immortalized Cells: How to Assess Immortality and Confirm Identity
Shitova, M., Alpeeva, E., & Vorotelyak, E. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms252313054 · Cited by 22
Metadata
- Authors
- Robertson, D. M., Li, L., Fisher, S., Pearce, V. P., Shay, J. W., Wright, W. E., Cavanagh, H. D., & Jester, J. V.
- Venue
- Investigative Opthalmology & Visual Science
- Published
- 2005-02-01
- Volume
- 46
- Issue
- 2
- Pages
- 470
- ISSN
- 0146-0404 (ISSN-L)
- 1552-5783
- OpenAlex
- W2030589581
- Cited by
- 286
- References
- 7
- FWCI
- 6.90
- Type
- journal article
- Language
- en
- Open access
- closed
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- Robertson, D. M., Li, L., Fisher, S., Pearce, V. P., Shay, J. W., Wright, W. E., Cavanagh, H. D., & Jester, J. V. (2005). Characterization of Growth and Differentiation in a Telomerase-Immortalized Human Corneal Epithelial Cell Line. Investigative Opthalmology & Visual Science. https://doi.org/10.1167/iovs.04-0528
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