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Telomerase-mediated immortalization of human vaginal wall fibroblasts derived from patients with pelvic organ prolapse

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Chinese Medical Journal 2023年第5期136卷 578-587页

作者： Tao Guo Ting Xie Jinghe Lang Zhijing Sun Department of Obstetrics and Gynecology Peking Union Medical College HospitalPeking Union Medical CollegeChinese Academy of Medical SciencesNational Clinical Research Center for Obstetric&Gynecologic DiseasesBeijing 100730China Department of Medical Research Center Peking Union Medical College HospitalChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100730China.

Background: Extracellular matrix (ECM) remodeling is the most important pathomechanism of pelvic organ prolapse (POP). Fibroblasts are the key to ECM regulation. The passaging capacity of human vaginal wall fibroblasts (hVWFs) is limited in vitro . Here, we aimed to immortalize hVWFs through the introduction of human telomerase reverse transcriptase (hTERT). Methods: Primary cells were derived from the vaginal wall tissue of patients with POP. Cellular senescence was detected via senescence-associated β-galactosidase staining. We employed a lentiviral transfection vector to stably express hTERT in hVWFs at passage 3, generating immortalized hVWFs (i-hVWFs). We then assessed cellular proliferation via the CCK-8 and EdU assays as well as cellular migration via wound healing assays. G-banded chromosome karyotypic analysis was performed to evaluate chromosomal karyotype stability. Finally, cellular tumorigenesis capacity was assessed in nude mice. A two-tailed Student’s t test was used to compare differences between the two groups. Results: Our results showed that senescence of primary hVWFs significantly increased from passage seven. From passage 11, hVWFs showed a significantly higher senescence percentage than i-hVWFs. During the continuous passage, i-hVWFs presented stability in proliferation, migration capacity, expression of ECM regulation-related genes, and chromosome karyotype. In vivo tumorigenesis was absent in i-hVWFs. Conclusions: The senescence of hVWFs significantly increased from the seventh passage, and we successfully used hTERT to immortalize hVWFs derived from patients with POP. Studies on POP that require a long-lived hVWF line will benefit from our technique.

关键词： Pelvic organ prolapse Fibroblasts telomere shortening Cellular senescence

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Role of mitophagy in the hallmarks of aging

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The Journal of Biomedical Research 2023年第1期37卷 1-14页

作者： Jie Wen Tingyu Pan Hongyan Li Haixia Fan Jinhua Liu Zhiyou Cai Bin Zhao Department and Institute of Neurology Guangdong Medical UniversityZhanjiangGuangdong 524001China Guangdong Key Laboratory of Aging-related Cardiac and Cerebral Diseases ZhanjiangGuangdong 524001China Chongqing Key Laboratory of Neurodegenerative Diseases Chongqing 400013China Department of Neurology Chongqing General HospitalChongqing 400013China Department of Neurology the Affiliated Hospital of Southwest Medical UniversityLuzhouSichuan 646000China Chongqing Medical University Chongqing 400042China

Aging, subjected to scientific scrutiny, is extensively defined as a time-dependent decline in functions that involves the majority of organisms. The time-dependent accretion of cellular lesions is generally a universal trigger of aging, while mitochondrial dysfunction is a sign of aging. Dysfunctional mitochondria are identified and removed by mitophagy, a selective form of macroautophagy. Increased mitochondrial damage resulting from reduced biogenesis and clearance may promote the aging process. The primary purpose of this paper is to illustrate in detail the effects of mitophagy on aging and emphasize the associations between mitophagy and other signs of aging, including dietary restriction, telomere shortening, epigenetic alterations, and protein imbalance.The evidence regarding the effects of these elements on aging is still limited. And although the understanding of relationship between mitophagy and aging has been long-awaited, to analyze details of such a relationship remains the main challenge in aging studies.

关键词： mitophagy aging dietary restriction telomere shortening epigenetic alterations protein imbalance

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Mesenchymal stem cells-based therapy as a potential treatment in neurodegenerative disorders: is the escape from senescence an answer?

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Neural Regeneration Research 2015年第6期10卷 850-858页

作者： Alessandro Castorina Marta Anna Szychlinska Rubina Marzagalli Giuseppe Musumeci Department of Biomedical and Biotechnological Science Section of Human Anatomy and HistologySchool of MedicineUniversity of Catania

Aging is the most prominent risk factor contributing to the development of neurodegenerative disorders. In the United States, over 35 million of elderly people suffer from age-related diseases. Aging impairs the self-repair ability of neuronal cells, which undergo progressive deterioration. Once initiated, this process hampers the already limited regenerative power of the central nervous system, making the search for new therapeutic strategies particularly difficult in elderly affected patients. So far, mesenchymal stem cells have proven to be a viable option to ameliorate certain aspects of neurodegeneration, as they possess high proliferative rate and differentiate in vitro into multiple lineages. However, accumulating data have demonstrated that during longterm culture, mesenchymal stem cells undergo spontaneous transformation. Transformed mesenchymal stem cells show typical features of senescence, including the progressive shortening of telomers, which results in cell loss and, as a consequence, hampered regenerative potential. These evidences, in line with those observed in mesenchymal stem cells isolated from old donors, suggest that senescence may represent a limit to mesenchymal stem cells exploitation in therapy, prompting scholars to either find alternative sources of pluripotent cells or to arrest the age-re- lated transformation. In the present review, we summarize findings from recent literature, and critically discuss some of the major hurdles encountered in the search of appropriate sources of mesenchymal stem cells, as well as benefits arising from their use in neurodegenerative diseases. Finally, we provide some insights that may aid in the development of strategies to arrest or, at least, delay the aging of mesenchymal stem cells to improve their therapeutic potential.

关键词： aging neurodegenerative disorders telomere shortening MSCs cellular therapy

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