Generation of a stable mouse endometrial tumor-derived cell line using conditional reprogramming.

Abstract

Background: Endometrial cancer (EC) is a common malignancy of the female reproductive tract worldwide. While comprehensive genomic analyses have identified multiple genetic alterations in EC, disease models remain relatively rare, limiting the ability to conduct disease studies. Establishing a hybrid in vitro and in vivo model of EC and comparing it to its in vivo counterpart could greatly advance research into EC mechanisms and treatment strategies.

Methods: We established genetically engineered EC mice (Pten ^flox/+; Stk11 ^flox/flox; LTF-Cre (PSC)) and constructed a corresponding epithelial cell line from isolated uterine tumor tissues using conditional reprogramming techniques. The cell line was successfully passaged, frozen, and stored in a biobank, providing a valuable resource for future EC research.

Results: The PSC-derived cell line stably retained the mutant alleles of Pten and Stk11 during passaging, preserving the genetic profile of the original tumor. In addition, we performed various functional assays on isolated mouse cell linesand formed tumors after subcutaneous implantation of cells in immunodeficient mice and C57BL/6 mice with an intact immune system. Hematoxylin and eosin (H&E) staining and immunohistochemical staining were used to characterize the histopathology and biomarkers of the subcutaneously implanted tumors, which were highly similar to those of the parental mouse uterine tumors.

Conclusions: The isolated mouse cell line provides a powerful platform for basic and translational EC studies, matching well with the corresponding in vivo models. The fidelity of the cell line to human-relevant genetic alterations and tumor biology makes it a valuable tool for investigating EC pathogenesis and evaluating novel therapeutic approaches.