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Longitudinal evaluation of primary uterine leiomyoma cultures reveals changes in molecular identity and drug responsiveness

Year: 2026

Type: article

Abstract: Uterine leiomyomas (ULs) are typically benign, yet they severely impair female fertility and reproductive health, representing a major unmet clinical need. While primary two-dimensional (2D) culture is widely utilized to study ULs, the longitudinal stability of these cells in standard environments remains poorly characterized. Herein, we systematically evaluated how experimental parameters and serial 2D passaging influence the molecular identity and drug responsiveness of patient-derived UL cells. Morphological, transcriptomic, and functional analyses revealed a rapid culture-induced loss of native tissue identity. Targeted gene profiling demonstrated that estrogen and progesterone receptor mRNA levels dropped precipitously by passage 2, followed by a progressive decline in the smooth muscle marker ACTA2 . Although optimized media transiently enhanced early hormone responsiveness, it failed to prevent long-term phenotypic drift. Furthermore, drug responses to selective progesterone receptor modulators (SPRMs) were highly context-dependent, significantly altered by prolonged culture and media composition. These results demonstrate that primary UL cells undergo rapid transcriptional reprogramming, leading to unstable pharmacological behavior. Consequently, defining a strictly restricted culture window at early passages establishes an indispensable methodological baseline to prevent artefactual data and ensure the reproducibility and translational fidelity of future in vitro pharmacological evaluations.

Source: Scientific Reports

DOI: 10.1038/s41598-026-69254-3