Optimized clonal isolation and immortalization of Rett syndrome patient fibroblasts for in vitro modeling of MECP2 mutations.

ABSTRACT: Rett syndrome (RTT) is a severe neurodevelopmental disorder primarily caused by mutations in the X-linked MECP2 gene. Patient-derived fibroblasts serve as a practical system to study systemic aspects of RTT, however, their limited proliferative capacity due to cellular senescence poses a significant challenge. In this study, we establish a robust workflow to isolate and immortalize clonal fibroblast lines from female RTT patients heterozygous for two distinct MECP2 mutations (c.705delG and 1155del32). By employing single-cell cloning prior to hTERT-mediated immortalization, we generated stable, proliferative fibroblast clones with verified clonality and severely skewed X-chromosome inactivation. Wildtype clones exclusively expressed full-length MeCP2 protein, whereas mutant clones exhibit

SUBMITTER: Sarne V 

PROVIDER: S-EPMC12518718 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature