Клеточное старение и технологии иммортализации в трансляционной медицине: от фундаментальных механизмов к терапевтическим платформам

Первичные клеточные культуры являются неотъемлемой частью фундаментальных исследований современной клеточной биологии. Их ценность обусловлена максимальным приближением к условиям in vivo, что повышает достоверность получаемых результатов. Однако широкое применение первичных клеток in vitro ограничено их естественным пролиферативным потенциалом – клеточным старением (сенесценцией), что создает сложности для проведения долгосрочных и воспроизводимых исследований. Сенесценция представляет собой сложный многокомпонентный ответ клетки на различные стимулы. В связи с этим выделяют репликативное и онкоген-индуцированное старение, а также преждевременное старение, вызванное стрессом. Каждый из этих типов сенесценции имеет уникальные признаки, изучение которых позволяет понять, как преодолеть пролиферативный барьер клетки путем иммортализации. Классические методы иммортализации, такие как экспрессия каталитической субъединицы теломеразы (TERT), использование вирусных онкогенов (SV40 Large T-антиген, HPV16 Е6/Е7) и протоонкогенов, несмотря на свою эффективность, имеют ряд ограничений, включая риск онкогенной трансформации и изменение исходного фенотипа клетки. В связи с этим набирают популярность современные методы условной (обратимой) иммортализации, использование неинтегрирующихся систем и комбинированных методов, которые позволяют сохранить высокий пролиферативный потенциал клеток, обеспечивая при этом максимальное сходство с первичными клетками. В данном обзоре литературы систематизированы современные представления о видах клеточной сенесценции. Подробно рассмотрены основные и вспомогательные методы иммортализации, их преимущества и недостатки. Дается характеристика современных иммортализованных клеточных линий с учетом их фенотипической стабильности и перспектив применения в регенеративной медицине, тканевой инженерии, 3D-культивировании и доклинической фармакологии. В работе проанализированы данные, преимущественно опубликованные в течение последних 15 лет.

1. Campisi J, d’Adda Di Fagagna F. Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol. 2007;8:729–40. doi:10.1038/nrm2233

2. Voloshin N, Tyurin-Kuzmin P, Karagyaur M, et al. Practical Use of Immortalized Cells in Medicine: Current Advances and Future Perspectives. IJMS. 2023;24:12716. doi:10.3390/ijms241612716

3. Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Experimental Cell Research. 1961;25:585–621. doi:10.1016/0014-4827(61)90192-6

4. Coppé JP, Desprez PY, Krtolica A, et al. The Senescence-Associated Secretory Phenotype: The Dark Side of Tumor Suppression. Annu Rev Pathol Mech Dis. 2010;5:99–118. doi:10.1146/annurev-pathol-121808-102144

5. Salama R, Sadaie M, Hoare M, et al. Cellular senescence and its effector programs. Genes Dev. 2014;28:99–114. doi:10.1101/gad.235184.113

6. Kumari R, Jat P. Mechanisms of Cellular Senescence: Cell Cycle Arrest and Senescence Associated Secretory Phenotype. Front Cell Dev Biol. 2021;9:645593. doi:10.3389/fcell.2021.645593

7. Nelson G, Wordsworth J, Wang C, et al. A senescent cell bystander effect: senescence‐induced senescence. Aging Cell. 2012;11:345–9. doi:10.1111/j.1474-9726.2012.00795.x

8. Wang B, Han J, Elisseeff JH, et al. The senescence-associated secretory phenotype and its physiological and pathological implications. Nat Rev Mol Cell Biol. 2024;25:958–78. doi:10.1038/s41580-024-00727-x

9. Prata LGPL, Ovsyannikova IG, Tchkonia T, et al. Senescent cell clearance by the immune system: Emerging therapeutic opportunities. Seminars in Immunology. 2018;40:101275. doi:10.1016/j.smim.2019.04.003

10. Ortiz-Montero P, Londoño-Vallejo A, Vernot JP. Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line. Cell Commun Signal. 2017;15:17. doi:10.1186/s12964-017-0172-3

11. Tominaga K, Suzuki HI. TGF-β Signaling in Cellular Senescence and Aging-Related Pathology. IJMS. 2019;20:5002. doi:10.3390/ijms20205002

12. Deng Z, Fan T, Xiao C, et al. TGF-β signaling in health, disease and therapeutics. Sig Transduct Target Ther. 2024;9:61. doi:10.1038/s41392-024-01764-w

13. Dong Z, Luo Y, Yuan Z, et al. Cellular senescence and SASP in tumor progression and therapeutic opportunities. Mol Cancer. 2024;23:181. doi:10.1186/s12943-024-02096-7

14. Robbins PD, Jurk D, Khosla S, et al. Senolytic Drugs: Reducing Senescent Cell Viability to Extend Health Span. Annu Rev Pharmacol Toxicol. 2021;61:779–803. doi:10.1146/annurev-pharmtox-050120-105018

15. Suda M, Paul KH, Tripathi U, et al. Targeting Cell Senescence and Senolytics: Novel Interventions for Age-Related Endocrine Dysfunction. Endocrine Reviews. 2024;45:655–75. doi:10.1210/endrev/bnae010

16. Ghadaouia S, Olivier MA, Martinez A, et al. Homologous recombination-mediated irreversible genome damage underlies telomere-induced senescence. Nucleic Acids Research. 2021;49:11690–707. doi:10.1093/nar/gkab965

17. Pizzul P, Rinaldi C, Bonetti D. The multistep path to replicative senescence onset: zooming on triggering and inhibitory events at telomeric DNA. Front Cell Dev Biol. 2023;11:1250264. doi:10.3389/fcell.2023.1250264

18. Zhu H, Blake S, Kusuma FK, et al. Oncogene-induced senescence: From biology to therapy. Mechanisms of Ageing and Development. 2020;187:111229. doi:10.1016/j.mad.2020.111229

19. Liu X ling, Ding J, Meng L hua. Oncogene-induced senescence: a double edged sword in cancer. Acta Pharmacol Sin. 2018;39:1553–8. doi:10.1038/aps.2017.198

20. Di Micco R, Sulli G, Dobreva M, et al. Interplay between oncogene-induced DNA damage response and heterochromatin in senescence and cancer. Nat Cell Biol. 2011;13:292–302. doi:10.1038/ncb2170

21. Toussaint O, Royer V, Salmon M, et al. Stress-induced premature senescence and tissue ageing. Biochemical Pharmacology. 2002;64:1007–9. doi:10.1016/S0006-2952(02)01170-X

22. Wiley CD, Velarde MC, Lecot P, et al. Mitochondrial Dysfunction Induces Senescence with a Distinct Secretory Phenotype. Cell Metabolism. 2016;23:303–14. doi:10.1016/j.cmet.2015.11.011

23. Van Der Zanden SY, Qiao X, Neefjes J. New insights into the activities and toxicities of the old anticancer drug doxorubicin. The FEBS Journal. 2021;288:6095–111. doi:10.1111/febs.15583

24. Bajtai E, Kiss C, Bakos É, et al. Therapy-induced senescence is a transient drug resistance mechanism in breast cancer. Mol Cancer. 2025;24:128. doi:10.1186/s12943-025-02310-0

25. Beltzig L, Schwarzenbach C, Leukel P, et al. Senescence Is the Main Trait Induced by Temozolomide in Glioblastoma Cells. Cancers. 2022;14:2233. doi:10.3390/cancers14092233

26. Schwarzenbach C, Rinke J, Vilar JB, et al. Therapy-induced senescence of glioblastoma cells is determined by the p21CIP1-CDK1/2 axis and does not require activation of DREAM. Cell Death Dis. 2025;16:357. doi:10.1038/s41419-025-07651-8

27. Zhong J, Chen J, Oyekan AA, et al. Ionizing Radiation Induces Disc Annulus Fibrosus Senescence and Matrix Catabolism via MMP-Mediated Pathways. IJMS. 2022;23:4014. doi:10.3390/ijms23074014

28. Yi Y, Xie H, Xiao X, et al. Ultraviolet A irradiation induces senescence in human dermal fibroblasts by down-regulating DNMT1 via ZEB1. Aging. 2018;10:212–28. doi:10.18632/aging.101383

29. Hernandez-Segura A, Nehme J, Demaria M. Hallmarks of Cellular Senescence. Trends in Cell Biology. 2018;28:436–53. doi:10.1016/j.tcb.2018.02.001

30. Sadaie M, Dillon C, Narita M, et al. Cell-based screen for altered nuclear phenotypes reveals senescence progression in polyploid cells after Aurora kinase B inhibition. Goldman RD, editor. MBoC. 2015;26:2971–85. doi:10.1091/mbc.E15-01-0003

31. Wallis R, Milligan D, Hughes B, et al. Senescence-associated morphological profiles (SAMPs): an image-based phenotypic profiling method for evaluating the inter and intra model heterogeneity of senescence. Aging. 2022;14:4220–46. doi:10.18632/aging.204072

32. Chalak M, Hesaraki M, Mirbahari SN, et al. Cell Immortality: In Vitro Effective Techniques to Achieve and Investigate Its Applications and Challenges. Life. 2024;14:417. doi:10.3390/life14030417

33. Lee KM, Choi KH, Ouellette MM. Use of exogenous hTERT to immortalize primary human cells. Cytotechnology. 2004;45:33–8. doi:10.1007/10.1007/s10616-004-5123-3

34. Guo T, Xie T, Lang J, et al. Telomerase-mediated immortalization of human vaginal wall fibroblasts derived from patients with pelvic organ prolapse. Chinese Medical Journal. 2023;136:578–87. doi:10.1097/CM9.0000000000002278

35. Farwell DG, Shera KA, Koop JI, et al. Genetic and Epigenetic Changes in Human Epithelial Cells Immortalized by Telomerase. The American Journal of Pathology. 2000;156:1537–47. doi:10.1016/S0002-9440(10)65025-0

36. Han Y, Li X, Zhang Y, et al. Mesenchymal Stem Cells for Regenerative Medicine. Cells. 2019;8:886. doi:10.3390/cells8080886

37. Hindle J, Williams A, Kim Y, et al. hTERT-Immortalized Mesenchymal Stem Cell-Derived Extracellular Vesicles: Large-Scale Manufacturing, Cargo Profiling, and Functional Effects in Retinal Epithelial Cells. Cells. 2024;13:861. doi:10.3390/cells13100861

38. Siska EK, Weisman I, Romano J, et al. Generation of an immortalized mesenchymal stem cell line producing a secreted biosensor protein for glucose monitoring. Shi XM, editor. PLoS ONE. 2017;12:e0185498. doi:10.1371/journal.pone.0185498

39. Denzer L, Muranyi W, Herold R, et al. Transcriptome and Functional Comparison of Primary and Immortalized Endothelial Cells of the Human Choroid Plexus at the Blood–Cerebrospinal Fluid Barrier. IJMS. 2025;26:1779. doi:10.3390/ijms26041779

40. Xie F, Gong K, Li K, et al. Reversible Immortalization Enables Seamless Transdifferentiation of Primary Fibroblasts into Other Lineage Cells. Stem Cells and Development. 2016;25:1243–8. doi:10.1089/scd.2016.0035

41. Cui H, Liang W, Wang D, et al. Establishment and Characterization of an Immortalized Porcine Oral Mucosal Epithelial Cell Line as a Cytopathogenic Model for Porcine Circovirus 2 Infection. Front Cell Infect Microbiol. 2019;9:171. doi:10.3389/fcimb.2019.00171

42. Liu J, Xu D, Yan J, et al. A novel H‑tert immortalized human sebaceous gland cell line (XL-i-20) for the investigation of photodynamic therapy. Photodiagnosis and Photodynamic Therapy. 2024;48:104238. doi:10.1016/j.pdpdt.2024.104238

43. Alyoshina NM, Ruchko ES, Tkachenko MD, et al. IMG-A1: A Novel Immortalized Granulosa Cell Line for Investigating FSH-Dependent Folliculogenesis and Ovarian Pathophysiology. Cells. 2025;14:1940. doi:10.3390/cells14241940

44. Kirsch-Volders M, Mišík M, Fenech M. Tetraploidy in normal tissues and diseases: mechanisms and consequences. Chromosoma. 2025;134:3. doi:10.1007/s00412-025-00829-1

45. Shenk EM, Ganem NJ. Generation and Purification of Tetraploid Cells. In: Chang P, Ohi R, editors. The Mitotic Spindle [Internet]. New York, NY: Springer New York; 2016 [cited 2026 Jun 16]. p. 393–401. (Methods in Molecular Biology). Available from: http://link.springer.com/10.1007/978-1-4939-3542-0_24 doi:10.1007/978-1-4939-3542-0_24

46. Alwin Prem Anand A, Gowri Sankar S, Kokila Vani V. Immortalization of neuronal progenitors using SV 40 large T antigen and differentiation towards dopaminergic neurons. J Cellular Molecular Medi. 2012;16:2592–610. doi:10.1111/j.1582-4934.2012.01607.x

47. Wang Y, Chen S, Yan Z, et al. A prospect of cell immortalization combined with matrix microenvironmental optimization strategy for tissue engineering and regeneration. Cell Biosci. 2019;9:7. doi:10.1186/s13578-018-0264-9

48. De Bardet JC, Cardentey CR, González BL, et al. Cell Immortalization: In Vivo Molecular Bases and In Vitro Techniques for Obtention. BioTech. 2023;12:14. doi:10.3390/biotech12010014

49. Dos Santos A, Lyu N, Balayan A, et al. Generation of Functional Immortalized Human Corneal Stromal Stem Cells. IJMS. 2022;23:13399. doi:10.3390/ijms232113399

50. Vafa O, Wade M, Kern S, et al. c-Myc Can Induce DNA Damage, Increase Reactive Oxygen Species, and Mitigate p53 Function. Molecular Cell. 2002;9:1031–44. doi:10.1016/S1097-2765(02)00520-8

51. McGrath KE, Olsen JL, Koniski AD, et al. BMI1 regulates human erythroid self-renewal through both gene repression and gene activation. Nat Commun. 2025;16:7619. doi:10.1038/s41467-025-62993-3

52. Dickson MA, Hahn WC, Ino Y, et al. Human Keratinocytes That Express hTERT and Also Bypass a p16INK4a -Enforced Mechanism That Limits Life Span Become Immortal yet Retain Normal Growth and Differentiation Characteristics. Molecular and Cellular Biology. 2000;20:1436–47. doi:10.1128/MCB.20.4.1436-1447.2000

53. Palomino Lago E, Roberts SJ, Orriss IR, et al. Generation and characterization of an immortalized equine osteoblast cell line. The Veterinary Journal. 2026;316:106573. doi:10.1016/j.tvjl.2026.106573

54. Faria J, Ahmed S, Stamatialis D, et al. Bioengineered Kidney Tubules Efficiently Clear Uremic Toxins in Experimental Dialysis Conditions. IJMS. 2023;24:12435. doi:10.3390/ijms241512435

55. Yamaguchi N, Horio E, Sonoda J, et al. Immortalization of Mesenchymal Stem Cells for Application in Regenerative Medicine and Their Potential Risks of Tumorigenesis. IJMS. 2024;25:13562. doi:10.3390/ijms252413562

56. Sutyagina OI, Beilin AK, Vorotelyak EA, et al. Immortalization Reversibility in the Context of Cell Therapy Biosafety. IJMS. 2023;24:7738. doi:10.3390/ijms24097738

57. Xu LW, Sun YD, Fu QY, et al. Unveiling senescence-associated secretory phenotype in epidermal aging: insights from reversibly immortalized keratinocytes. Aging. 2024;16:12651–66. doi:10.18632/aging.206117

58. Zhang C, Hao W, Yang Y, et al. Conditional immortalization of olfactory ensheathing cells for intelligently regulating their proliferation state. Sci Rep. 2025;15:30398. doi:10.1038/s41598-025-15144-z

59. Scharfmann R, Pechberty S, Hazhouz Y, et al. Development of a conditionally immortalized human pancreatic β cell line. J Clin Invest. 2014;124:2087–98. doi:10.1172/JCI72674

60. Papapetrou EP, Schambach A. Gene Insertion Into Genomic Safe Harbors for Human Gene Therapy. Molecular Therapy. 2016;24:678–84. doi:10.1038/mt.2016.38

61. Ordovás L, Boon R, Pistoni M, et al. Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition. Stem Cell Reports. 2018;10:673. doi:10.1016/j.stemcr.2018.01.034

62. Pellegrini S, Zamarian V, Sordi V. Strategies to Improve the Safety of iPSC-Derived β Cells for β Cell Replacement in Diabetes. Transpl Int. 2022;35:10575. doi:10.3389/ti.2022.10575

63. Qadir MMF, Álvarez-Cubela S, Belle K, et al. A Double Fail-Safe Approach to Prevent Tumorigenesis and Select Pancreatic β Cells from Human Embryonic Stem Cells. Stem Cell Reports. 2019;12:611–23. doi:10.1016/j.stemcr.2019.01.012

64. Fusaki N, Ban H, Nishiyama A, et al. Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc Jpn Acad, Ser B. 2009;85:348–62. doi:10.2183/pjab.85.348

65. Yu J, Hu K, Smuga-Otto K, et al. Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences. Science. 2009;324:797–801. doi:10.1126/science.1172482

66. Warren L, Manos PD, Ahfeldt T, et al. Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA. Cell Stem Cell. 2010;7:618–30. doi:10.1016/j.stem.2010.08.012

67. Oshimura M, Tabata T, Uno N, et al. Rejuvenation of human mesenchymal stem cells using a nonintegrative and conditionally removable Sendai virus vector. Sci Rep. 2024;14:23623. doi:10.1038/s41598-024-74757-y

68. Okawa M, Komatsu H, Kazuki Y, et al. Establishment of immortalized ovarian stromal cell lines using Sendai virus vectors: a platform for studying tumor–stroma interactions and carcinogenesis. Human Cell. 2025;39:15. doi:10.1007/s13577-025-01324-6

69. Ramunas J, Yakubov E, Brady JJ, et al. Transient delivery of modified mRNA encoding TERT rapidly extends telomeres in human cells. The FASEB Journal. 2015;29:1930–9. doi:10.1096/fj.14-259531

70. Cheng LC, You S, Ren T, et al. Development and characterization of a novel immortalized human vaginal fibroblast cell line for advanced applications in reproductive health. Reprod Biol Endocrinol. 2025;23:56. doi:10.1186/s12958-025-01393-0

71. Mansour F, Parisi L, Rihs S, et al. Immortalization of patient-derived lip cells for establishing 3D lip models. Front Cell Dev Biol. 2024;12:1449224. doi:10.3389/fcell.2024.1449224

72. Hayakawa T, Fujita F, Okada F, et al. Establishment and characterization of immortalized sweat gland myoepithelial cells. Sci Rep. 2022;12:7. doi:10.1038/s41598-021-03991-5

73. Green AJ, Anchang B, Akhtari FS, et al. Extending the Lymphoblastoid Cell Line Model for Drug Combination Pharmacogenomics. Pharmacogenomics. 2021;22:543–51. doi:10.2217/pgs-2020-0160

74. Zhong J, Wang H, Yang K, et al. Reversibly immortalized keratinocytes (iKera) facilitate re-epithelization and skin wound healing: Potential applications in cell-based skin tissue engineering. Bioactive Materials. 2022;9:523–40. doi:10.1016/j.bioactmat.2021.07.022

75. Li X, Zhang S, Zhang Z, et al. Development of immortalized Hertwig’s epithelial root sheath cell lines for cementum and dentin regeneration. Stem Cell Res Ther. 2019;10:3. doi:10.1186/s13287-018-1106-8

76. Trakarnsanga K, Griffiths RE, Wilson MC, et al. An immortalized adult human erythroid line facilitates sustainable and scalable generation of functional red cells. Nat Commun. 2017;8:14750. doi:10.1038/ncomms14750

77. Mu X, Sang Y, Fang C, et al. Immunotherapy of tumors with human telomerase reverse transcriptase immortalized human umbilical vein endothelial cells. International Journal of Oncology. 2015;47:1901–11. doi:10.3892/ijo.2015.3175

78. Amemori T, Romanyuk N, Jendelova P, et al. Human conditionally immortalized neural stem cells improve locomotor function after spinal cord injury in the rat. Stem Cell Res Ther. 2013;4:68. doi:10.1186/scrt219

79. Irfan Maqsood M, Matin MM, Bahrami AR, et al. Immortality of cell lines: challenges and advantages of establishment. Cell Biology International. 2013;37:1038–45. doi:10.1002/cbin.10137

80. Fan Y, Wong ST, Goh ELK, et al. Cell therapy for stroke and spinal cord injury in clinical trials. Stem Cells Translational Medicine. 2026;15:szaf082. doi:10.1093/stcltm/szaf082

81. Shitova M, Alpeeva E, Vorotelyak E. Review of hTERT-Immortalized Cells: How to Assess Immortality and Confirm Identity. IJMS. 2024;25:13054. doi:10.3390/ijms252313054

82. Cheng LC, You S, Ren T, et al. Development and characterization of a novel immortalized human vaginal fibroblast cell line for advanced applications in reproductive health. Reprod Biol Endocrinol. 2025;23:56. doi:10.1186/s12958-025-01393-0