Cellular senescence and immortalization technologies in translational medicine: from fundamental mechanisms to therapeutic platforms

Primary cell cultures are an indispensable tool in fundamental research within modern cell biology. Their value lies in their close physiological resemblance to in vivo conditions, which significantly enhances the reliability and translational relevance of experimental data. However, the broad application of primary cells in vitro is constrained by their finite proliferative capacity, culminating in cellular senescence, which poses substantial challenges for long-term and highly reproducible studies. Cellular senescence represents a complex, multifaceted cellular response to diverse stimuli. Consequently, it is broadly classified into replicative senescence, oncogene-induced senescence (OIS), and stress-induced premature senescence. Each of these senescence modalities exhibits distinct molecular signatures, and elucidating these features is crucial for understanding how to bypass the cellular proliferative barrier through immortalization. Classical immortalization strategies, such as the ectopic expression of the telomerase catalytic subunit (TERT) or the introduction of viral oncogenes (e.g., SV40 Large T-antigen, HPV16 E6/E7) and proto-oncogenes, are highly effective but carry significant limitations. These include the inherent risk of oncogenic transformation and the alteration of the native cellular phenotype. Consequently, contemporary approaches such as conditional immortalization, non-integrating delivery systems, and combinatorial strategies are gaining prominence. These methods aim to sustain a high proliferative potential while preserving maximal phenotypic fidelity to the original primary cells. This review systematizes current knowledge regarding the distinct modalities of cellular senescence. It provides a comprehensive analysis of both primary and auxiliary immortalization techniques, critically evaluating their respective advantages and limitations. Furthermore, it characterizes contemporary immortalized cell lines with a specific focus on their phenotypic stability and translational potential in regenerative medicine, tissue engineering, 3D cell culture models, and preclinical pharmacology. The analysis presented herein is primarily based on data published within the last 15 years.

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