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

作　　者：Tao Guo Ting Xie Jinghe Lang Zhijing Sun 

机构地区：[1]Department of Obstetrics and Gynecology,Peking Union Medical College Hospital,Peking Union Medical College,Chinese Academy of Medical Sciences,National Clinical Research Center for Obstetric&Gynecologic Diseases,Beijing 100730,China [2]Department of Medical Research Center,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China.

出　　处：《Chinese Medical Journal》2023年第5期578-587,共10页中华医学杂志（英文版）

基　　金：National Natural Science Foundation of China(No.81971366);Beijing Natural Science Foundation(No.Z190021);CAMS Innovation Fund for Medical Sciences(No.CIFMS 2020-I2M-C&T-B-043)

摘　　要：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 

分 类 号：R711.5[医药卫生—妇产科学]